Insect resistant plant

ABSTRACT

The present invention relates to novel pepper plants resistant to insects, and to seeds and fruits of said plants. The present invention also relates to methods of making and using such plants and their fruits. The invention further relates to markers and the use thereof in marker assisted breeding and for identifying the insect resistance trait. In particular, the present invention provides a cultivated  Capsicum annuum  plant which is resistant, particularly intermediately resistant, to infestations by insects of the family Thripidae and/or the genus  Bemisia , but especially to infestations by  Bemisia tabaci  and  Frankliniella occidentalis.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a division of U.S. application Ser. No. 12/451,236,which is a national phase application of International Application No.PCT/EP2008/055374, filed Apr. 30, 2008, which claims priority toEuropean Application No. 07290556.5, filed May 2, 2007 and EuropeanApplication No. 0711649.7, filed Oct. 30, 2007.

The present invention relates to novel pepper plants resistant toinsects, and to seeds and fruits of said plants. The present inventionalso relates to methods of making and using such plants and theirfruits. The invention further relates to markers and the use thereof inmarker assisted breeding and for identifying the insect resistancetrait.

Peppers are an important crop worldwide with an estimated commercialvalue of about 500 million dollars a year. Peppers are Solanaceas fromthe genus Capsicum, which includes the species Capsicum annuum, Capsicumfrutescens and Capsicum chinense. Commercial peppers are diploids withn=12 chromosomes. Peppers are cultivated and used around the world assweet peppers such as the bell pepper; or as pungent chili peppers,jalapeno peppers, and TABASCO® peppers; or as a source of dried powdersof various colors such as paprika. The types of cultivated peppers canbe differentiated by pungency, fruit shape, color and size (see forexample U.S. Pat. No. 6,498,287).

Pepper fruits, also commonly referred to as “peppers”, are highlyperishable. They are prone to water loss and shriveling, which rendersthem unappealing to customers. Pepper plants are also hosts to a numberof diseases. These diseases reduce the yield of the crops, but alsoaffect the appearance of the fruits, rendering them unmarketable. Inparticular, insects cause substantial crop damages, resulting insubstantial commercial losses. In some cases, the insects directlyaffect the plants or the fruits, in other cases they act as a vector forplants viruses. Usually insect damage reduces plant growth but does notcommonly kill the plant. Chemical control and crop rotation can be usedto reduce the damage caused by insects, but these strategies areexpensive and sometimes inconvenient.

Among insect pest affecting peppers, the white fly Bemisia tabaci(Hemiptera: Aleyrodidae) and various thrips species such as the WesternFlower Thrips: Frankfiniella occidentalis, the Onion Thrips: Thripstabaci, the Chilli Thrips Scirtothrips dorsalis, and the Melon ThripsThrips palmi are particularly devastating.

There are about 5000 described species of thrips (insects in the OrderThysanoptera). The species that feed on higher plants occur mostly inthe Family Thripidae. This family includes the important pest speciesincluding serious pests of ornamental, vegetable, and fruit crops in thefield and greenhouse. Feeding and egg-laying by thrips results indistortion, discoloration, silvering and bronzing of leaves and fruitsof vegetables reducing their market value. Some species of thrips arevectors of bunyaviruses (family Bunyaviridae, genus Tospovirus, typespecies tomato spotted wilt). Severe epidemics occur annually on food,fiber, and ornamental crops in tropical and subtropical regions of theworld.

The western flower thrips (Franklinella occidentalis) is anopportunistic insect pest in greenhouses which severely affects amultitude of crops. Frank occidentalis was spread nearly worldwide overthe past two decades. This thrips species is very damaging and difficultto control. It multiplies easily on pepper and creates physical damageson plant, flowers and fruits from the early stage of the nursery up tothe end of the crop. The larvae and adults feed on the epidermal cellsof leaves, buds, flowers and fruits. They affect the skin of the fruitand depreciate the marketable value. High-value greenhouse crops such asvegetables are particularly vulnerable to economic losses associatedwith thrips damage. Thrips is also an efficient vector of a devastatingvirus, the Tomato Spotted Wilt virus (TSWv) which creates big losses forthe growers. The infected plants present strong mosaic and necrosis onplants and fruits.

Thrips is difficult to control via chemical products as the insect hasdeveloped resistance to several insecticides used over the last 15years. Under greenhouse conditions, the use of biological predators,either with Onus in hot conditions or Amblyseius in cooler conditionsthat maintain a low level of thrips in the crop, is a wide spread butnot always a sufficient practice.

For the white fly, Bemisia tabaci, at least two biotypes have beendescribed: the B-type, identical to Bemisia argentifolii and the Q-type.

Control of Bemisia and thrips is particularly difficult, also because ofthe wide range of host plants. Bemisia and thrips species attack a widevariety of vegetable crops including tomato, beans, cucumbers, melons,bitter melon, capsicum, eggplant, pumpkin, squash and zucchini. Capsicumbelongs to the most seriously affected crops.

Because of the damages on plant and fruit and the transmission of adevastating virus, there is an unmet need for convenient andeconomically sustainable strategies to protect pepper crops againstthese pests. Host plant resistance is a good control strategy forBemisia and thrips. It is an environmentally friendly alternative forthe use of pesticides and may increase the efficiency of biologicalcontrol options and contribute to successful integrated pest managementprograms.

The present invention addresses this need by providing resistant pepperplants that are less attractive to insects and/or capable of resistinginsect infestation and/or development such as, for example, ovipositionand/or pupae development and would thus be to a considerable degreeprotected from insect infestations, particularly from infestations ofthe white fly Bemisia tabaci and/or thrips.

The present invention provides a cultivated Capsicum annuum plant whichis resistant, particularly intermediately resistant, to infestations byinsects of the family Thripidae and/or the genus Bemisia, but especiallyto infestations by Bemisia tabaci and Frankliniella occidentalis

Resistance to Bemisia infestations” or “Bemisia resistant plant” refersto the plants capability to resist attack, infestation, or colonizationby the insect. The level of resistance exhibited by a certain plant canbe scored, for example, by means of a standardized insect ResistanceAssay as described in Example 2A herein below using a scale from 1-9 forassessing the severity of the infestation.

In one embodiment, the invention provides a cultivated Capsicum annuumplant which is resistant, particularly intermediately resistant, toBemisia infestations, wherein said resistance can be assessed in astandard resistance assay, particularly an assay as described in Example2A below, and wherein a resistance score is obtained deviating by notmore than 3 scales, particularly by not more than 2 scales, butespecially by not more than 1 scale from a score obtainable with aCapsicum annuum plant of line 061M4387, representative seed of which isdeposited under Accession No, NUMB 41428, when assessed in the sameassay to a statistically significant extent and under identicalenvironmental conditions, particularly under the same insect pressure.

In one embodiment, a Bemisia resistant Capsicum annuum plant is providedthat is capable of resisting insect development, particularlyoviposition and/or pupae development on the plant such that the numberof pupae on the leaves of the plant determined in a standard resistanceassay, particularly an assay as described in Example 2A below, deviatesby not more than a factor of 20, particularly by not more than a factorof 15, more particularly by not more than a factor of 10, even moreparticularly by not more than a factor of 5, but especially by not morethan a factor of 2, from the number of pupae obtainable with a Capsicumannuum plant of line 061M4387, representative seed of which is depositedunder Accession No. NUMB 41428, when assessed in the same assay to astatistically significant extent and under identical environmentalconditions, particularly under the same insect pressure.

In one embodiment a Bemisia resistant Capsicum annuum plant is providedthat is capable of resisting insect development, particularlyoviposition and/or pupae development on the plant to essentially thesame extent as a Capsicum annuum plant of line 061M4387, representativeseed of which is deposited under Accession No. NCIMB 41428, whenassessed in the same assay to a statistically significant extent andunder identical environmental conditions, particularly under the sameinsect pressure.

In one embodiment, a Bemisia resistant Capsicum annuum plant is providedthat is capable of resisting insect development, particularlyoviposition and/or pupae development on the plant, wherein saidresistance can be assessed in a standard resistance assay, particularlyan assay as described in Example 2A below, and wherein a resistancescore is obtained that is at least 2 scales, particularly at least 3scales, more particularly at least 4 scales, but especially at least 5scales higher than the resistance score obtained with a standardsusceptible commercial variety, such as, for example, Vergasa orBikingo, when assessed in the same assay to a statistically significantextent and under identical environmental conditions, particularly underthe same insect pressure.

In one embodiment, a cultivated Capsicum annuum plant is provided, whichis resistant, particularly intermediately resistant, to thripsinfestations, especially to infestations with F. occidentalis,particularly by preventing plant damage caused by the feeding of thripson the epidermal cells of leaves, buds, flowers and fruits of theCapsicum annum plant, which results in silvering, loss of leaf colourand deformations of the developing fruit. The Capsicum plant'scapability of preventing feeding damage caused by thrips can be assessedin a standard resistance assay, particularly an assay as described inExample 2B below, by determining the extent of silvering damageaccording to a scale ranging from 1-9.

In one embodiment of the invention, a cultivated Capsicum annuum plantis provided, which is resistant, particularly intermediately resistant,to thrips infestations, particularly to infestations with F.occidentalis, particularly by preventing plant damage caused by thefeeding of thrips on the epidermal cells of leaves, buds, flowers andfruits of the Capsicum annuum plant, wherein said resistance can beassessed in a standard resistance assay, particularly an assay asdescribed in Example 2B below, and wherein a resistance score isobtained deviating by not more than 2 scales, particularly by not morethan 1 scale, but especially by not more than 0.5 scales from a scoreobtainable with a Capsicum annuum plant of line 061M4387, representativeseed of which is deposited under Accession No. NCIMB 41428, whenassessed in the same assay to a statistically significant extent andunder identical environmental conditions, particularly under the sameinsect pressure.

In one embodiment of the invention, the invention provides a cultivatedCapsicum annum plant which is resistant, particularly intermediatelyresistant, to thrips infestations, particularly to infestations with F.occidentalis, particularly by preventing plant damage caused by thefeeding of thrips on the epidermal cells of leaves, buds, flowers andfruits of the Capsicum annuum plant, wherein said resistance can beassessed in a standard resistance assay, particularly an assay asdescribed in Example 2B below, and wherein the silvering damage observeddoes not deviate by more than 8%, particularly by more than 5%, moreparticularly by more than 2%, even more particularly by more than 1%,but especially by more than 0.5%, from the damage exhibited on aCapsicum annuum plant of line 061M4387, representative seed of which isdeposited under Accession No. NCIMB 41428, when assessed in the sameassay to a statistically significant extent and under identicalenvironmental conditions, particularly under the same insect pressure.

In one embodiment of the invention, the invention provides a cultivatedCapsicum annum plant which is resistant, particularly intermediatelyresistant, to thrips infestations, particularly to infestations with F.occidentalis, particularly by preventing plant damage caused by thefeeding of thrips on the epidermal cells of leaves, buds, flowers andfruits of the Capsicum annuum plant, to essentially the same extent as aCapsicum annuum plant of line 061M4387, representative seed of which isdeposited under Accession No. NCIMB 41428, when assessed in the sameassay to a statistically significant extent and under identicalenvironmental conditions, particularly under the same insect pressure.

In one embodiment, a cultivated Capsicum annuum plant is provided, whichis resistant, particularly intermediately resistant, to Bemisia andthrips infestations, especially to infestations with Bemisia tabaci andF. occidentalis, particularly by preventing oviposition and/or pupaedevelopment of Bemisia and by preventing plant damage caused by thefeeding of thrips on the epidermal cells of leaves, buds, flowers andfruits of the Capsicum annuum plant, respectively, wherein saidresistance can be assessed in a standard resistance assay, particularlyan assay as described in Examples 2A and 2B below, and wherein, forBemisia, a resistance score is obtained deviating by not more than 3scales, particularly by not more than 2 scales, but especially by notmore than 1 scale from a score obtainable with a Capsicum annuum plantof line 061M4387, representative seed of which is deposited underAccession No. NCIMB 41428, and, for thrips, a resistance score isobtained deviating by not more than 2 scales, particularly by not morethan 1 scale, but especially by not more than 0.5 scales from a scoreobtainable with a Capsicum annuum plant of line 061M4387, representativeseed of which is deposited under Accession No. NCIMB 41428, whenassessed in the same assay to a statistically significant extent andunder identical environmental conditions, particularly under the sameinsect pressure.

In one embodiment, a cultivated Capsicum annuum plant is provided, whichis resistant, particularly intermediately resistant, to Bemisia andthrips infestations, especially to infestations with Bemisia tabaci andF. occidentalis, particularly by preventing oviposition and/or pupaedevelopment of Bemisia and by preventing plant damage caused by thefeeding of thrips on the epidermal cells of leaves, buds, flowers andfruits of the Capsicum annuum plant, respectively, to essentially thesame extent as a Capsicum annuum plant of line 061M4387, representativeseed of which is deposited under Accession No, NCIMB 41428, whenassessed in the same assay to a statistically significant extent andunder identical environmental conditions, particularly under the sameinsect pressure.

In one embodiment, the present invention provides a cultivated Capsicumannuum plant which is resistant, particularly intermediately resistant,to Bemisia infestations, wherein said plant contains a genome comprisingat least one quantitative trait focus (“QTL”) which contributes toBemisia resistance, in particular a cultivated Capsicum annum plantwhich is resistant, particularly intermediately resistant, to Bemisiainfestations, wherein said plant contains a genome comprising aquantitative trait locus (“QTL”) which contributes to Bemisiaresistance, wherein said QTL is located on chromosome 3 and/orchromosome 5.

In one embodiment, the present invention provides a cultivated Capsicumannuum plant which is resistant, particularly intermediately resistant,to Bemisia infestations, wherein said plant contains a genome comprisingat least two quantitative trait loci (“QTL”) which contribute to Bemisiaresistance, wherein a first QTL is located on chromosome 3 and an secondQTL is located on chromosome 5.

In one embodiment, the present invention provides a cultivated Capsicumannuum plant which is resistant, particularly intermediately resistant,to thrips infestations, wherein said plant contains a genome comprisingat least one quantitative trait locus (“QTL”) which contributes tothrips resistance, in particular a cultivated Capsicum annuum plantwhich is resistant, particularly intermediately resistant, to thripsinfestations, wherein said plant contains a genome comprising aquantitative trait locus (“QTL”) which contributes to thrips resistance,wherein said QTL is located on chromosome 5.

In one embodiment, the present invention provides a cultivated Capsicumannuum plant which is resistant, particularly intermediately resistant,to Bemisia and thrips infestations, wherein said plant contains a genomecomprising at least one quantitative trait locus (“QTL”) whichcontributes to Bemisia resistance and at least one quantitative traitlocus (“QTL”) which contributes to thrips resistance, respectively. Inparticular, the invention relates to a cultivated Capsicum annuum plantwhich is resistant, particularly intermediately resistant, to Bemisiaand thrips infestations as described herein before, wherein said QTLcontributing to Bemisia resistance is located on chromosome 3 and/orchromosome 5 and said QTL contributing to thrips resistance is locatedon chromosome 5.

In one embodiment, a cultivated Capsicum annuum plant is provided whichis resistant, particularly intermediately resistant, to Bemisia andthrips infestations, wherein said plant contains a genome comprisingquantitative trait loci (“QTL”) which contribute to Bemisia and thripsresistance, wherein a first QTL contributing to Bemisia resistance islocated on chromosome 3 and an second QTL contributing to Bemisiaresistance is located on chromosome 5, and said QTL contributing tothrips resistance is located on chromosome 5.

In one embodiment, the QTL on chromosome 5 is a single QTL contributingto both Bemisia and thrips resistance.

In one embodiment, said QTL are obtainable from a plant which has thegenetic background of line 061M4387, particularly from a plant which hasthe genetic background or architecture at the QTL of line 061M4387, butespecially from a plant of line 061M4387, representative seed of whichis deposited at NCIMB under Accession No. NCIMB 41428, or from a progenyor an ancestor thereof comprising said QTL.

In a further embodiment, the invention relates to a cultivated Capsicumannuum plant according to the invention and as described herein before,which plant contains a genome comprising at least one quantitative traitlocus (“QTL”) which contributes to Bemisia resistance, wherein said QTLis characterized by being genetically linked to at least one markerlocus, particularly to at least two marker loci, more particularly to atleast three marker loci and even more particularly to at least fourmarker loci, but especially to at least five and up to six marker loci,which marker loci are on chromosome 3 and co-segregate with the Bemisiaresistance trait and can be identified by a pair of PCR oligonucleotideprimers selected from the group of primer pair 1 represented by aforward primer of SEQ ID NO: 1 and a reverse primer of SEQ ID NO: 2,identifying marker locus 1; primer pair 2 represented by a forwardprimer of SEQ ID NO: 3 and a reverse primer of SEQ ID NO: 4, identifyingmarker locus 2; primer pair 3 represented by a forward primer of SEQ IDNO: 5 and a reverse primer of SEQ ID NO: 6, identifying marker locus 3;primer pair 4 represented by a forward primer of SEQ ID NO: 7 and areverse primer of SEQ ID NO: 8, identifying marker locus 4; primer pair5 represented by a forward primer of SEQ ID NO: 9 and a reverse primerof SEQ ID NO: 10, identifying marker locus 5; and primer pair 6represented by a forward primer of SEQ ID NO: 11 and a reverse primer ofSEQ ID NO: 12, identifying marker locus 6; or by any other marker locusthat is statistically correlated to the Bemisia resistance trait.

In one embodiment, the invention relates to a cultivated Capsicum annuumplant containing a genome comprising at least one quantitative traitlocus (“QTL”) which contributes to Bemisia resistance, wherein said QTLis obtainable from a donor plant which has the genetic background ofline 061M4387, particularly from a plant which has the geneticbackground or architecture at the QTL of line 061M4387, but especiallyfrom a plant of line 061M4387, representative seed of which is depositedat NCIMB under Accession No. NCIMB 41428, or from a progeny or anancestor thereof comprising said QTL, which QTL in the donor plant isgenetically linked to at least one marker locus, particularly to atleast two marker loci, particularly to at least three marker loci andparticularly to at least four marker loci, particularly to at least fivemarker loci, particularly to at least six marker loci, and up to sevenmarker loci, which marker loci are on chromosome 3 and co-segregate withthe Bemisia resistance trait and can be identified by a pair of PCRoligonucleotide primers selected from the group of primer pairs 1 to 6as given in SEQ ID NOs: 1 to 12.

In one embodiment, the invention relates to a cultivated Capsicum annuumplant according to the invention and as described herein before, whichplant contains a genome comprising a quantitative trait locus (“QTL”)which contributes to thrips resistance, wherein said QTL ischaracterized by being genetically linked to at least one marker locus,particularly to at least two marker loci, particularly to at least threemarker loci and particularly to at least four marker loci, particularlyto at least five marker loci particularly to at least six marker loci,and up to seven marker loci, which marker loci are on chromosome 5 andco-segregate with the Bemisia resistance trait and can be identified bya pair of PCR oligonucleotide primers selected from the group of primerpair 7 represented by a forward primer of SEQ ID NO: 13 and a reverseprimer of SEQ ID NO: 14, identifying marker locus 7; primer pair 8represented by a forward primer of SEQ ID NO: 15 and a reverse primer ofSEQ ID NO: 16, identifying marker locus 8; primer pair 9 represented bya forward primer of SEQ ID NO: 17 and a reverse primer of SEQ ID NO: 18,identifying marker locus 9; primer pair 10 represented by a forwardprimer of SEQ ID NO: 19 and a reverse primer of SEQ ID NO: 20,identifying marker locus 10; primer pair 11 represented by a forwardprimer of SEQ ID NO: 21 and a reverse primer of SEQ ID NO: 22,identifying marker locus 11; primer pair 12 represented by a forwardprimer of SEQ ID NO: 23 and a reverse primer of SEQ ID NO: 24,identifying marker locus 12, and primer pair 13 represented by a forwardprimer of SEQ ID NO: 25 and a reverse primer of SEQ ID NO: 26,identifying marker locus 13; or by any other marker locus that isstatistically correlated to the Bemisia resistance trait.

In one embodiment, the invention relates to a cultivated Capsicum annuumplant containing a genome comprising at least one quantitative traitlocus (“QTL”) which contributes to Bemisia resistance, wherein said QTLis obtainable from a donor plant which has the genetic background ofline 061M4387, particularly from a plant which has the geneticbackground or architecture at the QTL of line 061M4387, but especiallyfrom a plant of line 061M4387, representative seed of which is depositedat NCIMB under Accession No. NCIMB 41428, or from a progeny or anancestor thereof comprising said QTL, which QTL in the donor plant isgenetically linked to at least one marker locus, particularly to atleast two marker loci, particularly to at least three marker loci andparticularly to at least four marker loci, particularly to at least fivemarker loci, particularly to at least six marker loci, and up to sevenmarker loci, which marker loci are on chromosome 5 and co-segregate withthe Bemisia resistance trait and can be identified by a pair of PCRoligonucleotide primers selected from the group of primer pairs 7 to 13as given in SEQ ID NOs: 13 to 26.

In a further embodiment, the invention relates to a cultivated Capsicumannuum plant according to the invention and as described herein before,which plant contains a genome comprising at least two quantitative traitloci (“QTL”) which contribute to Bemisia resistance, wherein

-   -   a) a first QTL is characterized by being genetically linked to        at least one marker locus, particularly to at least two marker        loci, more particularly to at least three marker loci and even        more particularly to at least four marker loci, but especially        to at least five and up to six marker loci, which marker loci        are on chromosome 3 and co-segregate with the Bemisia resistance        trait and can be identified by a pair of PCR oligonucleotide        primers selected from the group of primer pair 1 represented by        a forward primer of SEQ ID NO: 1 and a reverse primer of SEQ ID        NO: 2, identifying marker locus 1; primer pair 2 represented by        a forward primer of SEQ ID NO: 3 and a reverse primer of SEQ ID        NO: 4, identifying marker locus 2; primer pair 3 represented by        a forward primer of SEQ ID NO: 5 and a reverse primer of SEQ ID        NO: 6, identifying marker locus 3; primer pair 4 represented by        a forward primer of SEQ ID NO: 7 and a reverse primer of SEQ ID        NO: 8, identifying marker locus 4: primer pair 5 represented by        a forward primer of SEQ ID NO: 9 and a reverse primer of SEQ ID        NO: 10, identifying marker locus 5; and primer pair 6        represented by a forward primer of SEQ ID NO: 11 and a reverse        primer of SEQ ID NO: 12, identifying marker locus 6; or by any        other marker locus on chromosome 3 that is statistically        correlated to the Bemisia resistance trait; and    -   b) a second QTL is characterized by being genetically linked to        at least one marker locus, particularly to at least two marker        loci, particularly to at least three marker loci and        particularly to at least four marker loci, particularly to at        least five marker loci, particularly to at least six marker        loci, and up to seven marker loci, which marker loci are on        chromosome 5 and co-segregate with the Bemisia resistance trait        and can be identified by a pair of PCR oligonucleotide primers        selected from the group of primer pair 7 represented by a        forward primer of SEQ ID NO: 13 and a reverse primer of SEQ ID        NO: 14, identifying marker locus 7; primer pair 8 represented by        a forward primer of SEQ ID NO: 15 and a reverse primer of SEQ ID        NO: 16, identifying marker locus 8; primer pair 9 represented by        a forward primer of SEQ ID NO: 17 and a reverse primer of SEQ ID        NO: 18, identifying marker locus 9; primer pair 10 represented        by a forward primer of SEQ ID NO: 19 and a reverse primer of SEQ        ID NO: 20, identifying marker locus 10; primer pair 11        represented by a forward primer of SEQ ID NO: 21 and a reverse        primer of SEQ ID NO: 22, identifying marker locus 11; primer        pair 12 represented by a forward primer of SEQ ID NO: 23 and a        reverse primer of SEQ ID NO: 24, identifying marker locus 12,        and primer pair 13 represented by a forward primer of SEQ ID NO:        25 and a reverse primer of SEQ ID NO: 26, identifying marker        locus 13, or by any other marker locus on chromosome 5 that is        statistically correlated to the Bemisia resistance trait.

In one embodiment, the invention relates to a cultivated Capsicum annuumplant containing a genome comprising at least two quantitative traitloci (“QTL”) which contribute to Bemisia resistance, wherein said QTLare obtainable from a donor plant which has the genetic background ofline 061M4387, particularly from a plant which has the geneticbackground or architecture at the QTL of line 061M4387, but especiallyfrom a plant of line 061M4387, representative seed of which is depositedat NCIMB under Accession No. NCIMB 41428, or from a progeny or anancestor thereof comprising said QTL, which first QTL is located onchromosome 3 in the donor plant and genetically linked to at least onemarker locus, particularly to at least two marker loci, particularly toat least three marker loci and particularly to at least four marker lad,particularly to at least five marker loci, particularly to at least sixmarker loci, which marker loci are on chromosome 3 and co-segregate withthe Bemisia resistance trait and can be identified by a pair of PCRoligonucleotide primers 1 to 6 as given in SEQ ID NOs: 1 to 12 and whichsecond QTL is located on chromosome 5 in the donor plant and geneticallylinked to at least one marker locus, particularly to at least two markerloci, particularly to at least three marker loci and particularly to atleast four marker loci, particularly to at least five marker loci,particularly to at least six marker loci, and up to seven marker loci,which marker loci are on chromosome 5 and co-segregate with the Bemisiaresistance trait and can be identified by a pair of PCR oligonucleotideprimers selected from the group of primer pairs 7 to 13 as given in SEQID NOs: 13 to 26.

In one embodiment, the invention relates to a cultivated Capsicum annuumplant according to the invention and as described herein before, whichplant contains a genome comprising a quantitative trait locus (“QTL”)which contributes to thrips resistance, wherein said QTL ischaracterized by being genetically linked to at least one marker locus,particularly to at least two marker loci, particularly to at least threemarker loci and particularly to at least four marker loci, particularlyto at least five marker loci, particularly to at least six marker loci,and up to seven marker loci, which marker loci are on chromosome 5 andco-segregate with the thrips resistance trait and can be identified by apair of PCR oligonucleotide primers selected from the group of primerpair 7 represented by a forward primer of SEQ ID NO: 13 and a reverseprimer of SEQ ID NO: 14, identifying marker locus 7; primer pair 8represented by a forward primer of SEQ ID NO: 15 and a reverse primer ofSEQ ID NO: 16, identifying marker locus 8; primer pair 9 represented bya forward primer of SEQ ID NO: 17 and a reverse primer of SEQ ID NO: 18,identifying marker locus 9; primer pair 10 represented by a forwardprimer of SEQ ID NO: 19 and a reverse primer of SEQ ID NO: 20,identifying marker locus 10; primer pair 11 represented by a forwardprimer of SEQ ID NO: 21 and a reverse primer of SEQ ID NO: 22,identifying marker locus 11; primer pair 12 represented by a forwardprimer of SEQ ID NO: 23 and a reverse primer of SEQ ID NO: 24,identifying marker locus 12, and primer pair 13 represented by a forwardprimer of SEQ ID NO: 25 and a reverse primer of SEQ ID NO: 26,identifying marker locus 13, or by any other marker locus on chromosome5 that is statistically correlated to the thrips resistance trait.

In one embodiment, the invention relates to a cultivated Capsicum annuumplant containing a genome comprising at least one quantitative traitlocus (“QTL”) which contributes to thrips resistance, wherein said QTLis obtainable from a donor plant which has the genetic background ofline 061M4387, particularly from a plant which has the geneticbackground or architecture at the QTL of line 061M4387, but especiallyfrom a plant of line 061M4387, representative seed of which is depositedat NCIMB under Accession No. NCIMB 41428, or from a progeny or anancestor thereof comprising said QTL, which QTL in the donor plant isgenetically linked to at least one marker locus, particularly to atleast two marker loci, particularly to at least three marker loci andparticularly to at least four marker loci, particularly to at least fivemarker loci, particularly to at least six marker loci, and up to sevenmarker loci, which marker loci are on chromosome 5 and co-segregate withthe thrips resistance trait and can be identified by a pair of PCRoligonucleotide primers selected from the group of primer pairs 7 to 13as given in SEQ ID NOs: 13 to 26.

In one embodiment, the present invention provides a cultivated Capsicumannuum plant which is resistant, particularly intermediately resistant,to Bemisia and thrips infestations, wherein said plant contains a genomecomprising at least one quantitative trait locus (“QTL”) whichcontributes to Bemisia resistance and at least one quantitative traitlocus (“QTL”) which contributes to thrips resistance, respectively,wherein said QTL contributing to

-   -   a) Bemisia resistance is characterized by being genetically        linked to at least one marker locus, particularly to at least        two marker loci, more particularly to at least three marker loci        and even more particularly to at least four marker loci, but        especially to at least five and up to six marker loci, which        marker loci are on chromosome 3 and co-segregate with the        Bemisia resistance trait and can be identified by a pair of PCR        oligonucleotide primers selected from the group of primer pair 1        represented by a forward primer of SEQ ID NO: 1 and a reverse        primer of SEQ ID NO: 2, identifying marker locus 1; primer pair        2 represented by a forward primer of SEQ ID NO: 3 and a reverse        primer of SEQ ID NO: 4, identifying marker locus 2; primer pair        3 represented by a forward primer of SEQ ID NO: 5 and a reverse        primer of SEQ ID NO: 6, identifying marker locus 3; primer pair        4 represented by a forward primer of SEQ ID NO: 7 and a reverse        primer of SEQ ID NO: 8, identifying marker locus 4; primer pair        5 represented by a forward primer of SEQ ID NO: 9 and a reverse        primer of SEQ ID NO: 10, identifying marker locus 5; and primer        pair 6 represented by a forward primer of SEQ ID NO: 11 and a        reverse primer of SEQ ID NO: 12, identifying marker locus 6, or        by any other marker locus on chromosome 3 that is statistically        correlated to the Bemisia resistance trait; and    -   b) thrips resistance is characterized by being genetically        linked to at least one marker locus, particularly to at least        two marker loci, particularly to at least three marker loci and        particularly to at least four marker loci, particularly to at        least five marker loci, particularly to at least six marker        loci, and up to seven marker loci, which marker loci are on        chromosome 5 and co-segregate with the thrips resistance trait        and can be identified by a pair of PCR oligonucleotide primers        selected from the group of primer pair 7 represented by a        forward primer of SEQ ID NO: 13 and a reverse primer of SEQ ID        NO: 14, identifying marker locus 7; primer pair 8 represented by        a forward primer of SEQ ID NO: 15 and a reverse primer of SEQ ID        NO; 16, identifying marker locus 8; primer pair 9 represented by        a forward primer of SEQ ID NO: 17 and a reverse primer of SEQ ID        NO: 18, identifying marker locus 9; primer pair 10 represented        by a forward primer of SEQ ID NO: 19 and a reverse primer of SEQ        ID NO: 20, identifying marker locus 10; primer pair 11        represented by a forward primer of SEQ ID NO: 21 and a reverse        primer of SEQ ID NO: 22, identifying marker locus 11; primer        pair 12 represented by a forward primer of SEQ ID NO: 23 and a        reverse primer of SEQ ID NO: 24, identifying marker locus 12,        and primer pair 13 represented by a forward primer of SEQ ID NO:        25 and a reverse primer of SEQ ID NO: 26, identifying marker        locus 13, or by any other marker locus on chromosome 5 that is        statistically correlated to the thrips resistance trait.

In one embodiment, the present invention provides a cultivated Capsicumannuum plant according to any of the preceding claims, which isresistant, particularly intermediately resistant, to Bemisia and thripsinfestations, wherein said plant contains a genome comprising at leastone quantitative trait locus (“QTL”) which contributes to Bemisiaresistance and at least one quantitative trait locus (“QTL”) whichcontributes to thrips resistance, respectively, wherein said QTLcontributing to

-   -   a) Bemisia resistance is characterized by being genetically        linked to        -   i. at least one marker locus, particularly to at least two            marker loci, more particularly to at least three marker loci            and even more particularly to at least four marker loci, but            especially to at least five and up to six marker loci, which            marker loci are on chromosome 3 and co-segregate with the            Bemisia resistance trait and can be identified by a pair of            PCR oligonucleotide primers selected from the group of            primer pair 1 represented by a forward primer of SEQ ID NO:            1 and a reverse primer of SEQ ID NO: 2, identifying marker            locus 1; primer pair 2 represented by a forward primer of            SEQ ID NO: 3 and a reverse primer of SEQ ID NO: 4,            identifying marker locus 2; primer pair 3 represented by a            forward primer of SEQ ID NO: 5 and a reverse primer of SEQ            ID NO: 6, identifying marker locus 3; primer pair 4            represented by a forward primer of SEQ ID NO: 7 and a            reverse primer of SEQ ID NO: 8, identifying marker locus 4;            primer pair 5 represented by a forward primer of SEQ ID NO:            9 and a reverse primer of SEQ ID NO: 10, identifying marker            locus 5; and primer pair 6 represented by a forward primer            of SEQ ID NO: 11 and a reverse primer of SEQ ID NO: 12,            identifying marker locus 6, or by any other marker locus on            chromosome 3 that is statistically correlated to the Bemisia            resistance trait; and/or        -   ii. at least one marker locus, particularly to at least two            marker loci, particularly to at least three marker loci and            particularly to at least four marker loci, particularly to            at least five marker loci, particularly to at least six            marker loci, and up to seven marker loci, which marker loci            are on chromosome 5 and co-segregate with the thrips            resistance trait and can be identified by a pair of PCR            oligonucleotide primers selected from the group of primer            pair 7 represented by a forward primer of SEQ ID NO: 13 and            a reverse primer of SEQ ID NO: 14, identifying marker locus            7; primer pair 8 represented by a forward primer of SEQ ID            NO: 15 and a reverse primer of SEQ ID NO: 16, identifying            marker locus 8; primer pair 9 represented by a forward            primer of SEQ ID NO: 17 and a reverse primer of SEQ ID NO:            18, identifying marker locus 9; primer pair 10 represented            by a forward primer of SEQ ID NO: 19 and a reverse primer of            SEQ ID NO: 20, identifying marker locus 10; primer pair 11            represented by a forward primer of SEQ ID NO: 21 and a            reverse primer of SEQ ID NO: 22, identifying marker locus            11; primer pair 12 represented by a forward primer of SEQ ID            NO: 23 and a reverse primer of SEQ ID NO: 24, identifying            marker locus 12, and primer pair 13 represented by a forward            primer of SEQ ID NO: 25 and a reverse primer of SEQ ID NO:            26, identifying marker locus 13, or by any other marker            locus on chromosome 5 that is statistically correlated to            the Bemisia resistance trait; and        -   b) thrips resistance is characterized by being genetically            linked to at least one marker locus, particularly to at            least two marker loci, particularly to at least three marker            loci and particularly to at least four marker loci,            particularly to at least five marker loci, particularly to            at least six marker loci, and up to seven marker loci, which            marker loci are on chromosome 5 and co-segregate with the            thrips resistance trait and can be identified by a pair of            PCR oligonucleotide primers selected from the group of            primer pair 7 represented by a forward primer of SEQ ID NO:            13 and a reverse primer of SEQ ID NO: 14, identifying marker            locus 7; primer pair 8 represented by a forward primer of            SEQ ID NO: 15 and a reverse primer of SEQ ID NO: 16,            identifying marker locus 8; primer pair 9 represented by a            forward primer of SEQ ID NO: 17 and a reverse primer of SEQ            ID NO: 18, identifying marker locus 9; primer pair 10            represented by a forward primer of SEQ ID NO: 19 and a            reverse primer of SEQ ID NO: 20, identifying marker locus            10; primer pair 11 represented by a forward primer of SEQ ID            NO: 21 and a reverse primer of SEQ ID NO: 22, identifying            marker locus 11; primer pair 12 represented by a forward            primer of SEQ ID NO: 23 and a reverse primer of SEQ ID NO:            24, identifying marker locus 12, and primer pair 13            represented by a forward primer of SEQ ID NO: 25 and a            reverse primer of SEQ ID NO: 26, identifying marker locus            13, or by any other marker locus on chromosome 5 that is            statistically correlated to the thrips resistance trait.

In one embodiment, the present invention provides a cultivated Capsicumannuum plant which is resistant, particularly intermediately resistant,to Bemisia and thrips infestations, wherein said plant contains a genomecomprising at least one quantitative trait locus (“QTL”) whichcontributes to Bemisia resistance and at least one quantitative traitlocus (“QTL”) which contributes to thrips resistance, respectively,wherein said QTL are obtainable from a donor plant which has the geneticbackground of line 061M4387, particularly from a plant which has thegenetic background or architecture at the QTL of line 061M4387, butespecially from a plant of line 061M4387, representative seed of whichis deposited at NCIMB under Accession No. NCIMB 41428, or from a progenyor an ancestor thereof comprising said QTL, and wherein

-   -   a) a first QTL contributing to Bemisia resistance is genetically        linked in the donor plant to at least one marker locus,        particularly to at least two marker loci, more particularly to        at least three marker loci and even more particularly to at        least four marker loci, but especially to at least five and up        to six marker loci, which marker loci are on chromosome 3 and        co-segregate with the Bemisia resistance trait and can be        identified by a pair of PCR oligonucleotide primers selected        from the group of primer pairs 1 to 6 represented by a forward        and a reverse primer as given in SEQ ID NOs: 1 to 12; and/or    -   b) a second QTL contributing to Bemisia resistance is        genetically linked in the donor plant to at least one marker        locus, particularly to at least two marker loci, particularly to        at least three marker loci and particularly to at least four        marker loci, particularly to at least five marker loci,        particularly to at least six marker loci, and up to seven marker        loci, which marker loci are on chromosome 5 and co-segregate        with the Bemisia resistance trait and can be identified by a        pair of PCR oligonucleotide primers selected from the group of        primer pairs 7 to 13 represented by a forward and a reverse        primer as given in SEQ ID NOs: 13 to 26; and.    -   c) a QTL contributing to thrips resistance is genetically linked        in the donor plant to at least one marker locus, particularly to        at least two marker loci, particularly to at least three marker        loci and particularly to at least four marker loci, particularly        to at least five marker loci, particularly to at least six        marker loci, and up to seven marker loci, which marker loci are        on chromosome 5 and co-segregate with the thrips resistance        trait and can be identified by a pair of PCR oligonucleotide        primers selected from the group of primer pairs 7 to 13        represented by a forward and a reverse primer as given in SEQ ID        NOs: 13 to 26.

In one embodiment, the present invention provides a cultivated Capsicumannuum plant which is resistant, particularly intermediately resistant,to Bemisia and thrips infestations, wherein said plant contains a genomecomprising at least one quantitative trait locus (“QTL”) whichcontributes to Bemisia resistance and at least one quantitative traitlocus (“QTL”) which contributes to thrips resistance, respectively,wherein said QTL are obtainable from a donor plant which has the geneticbackground of line 061M4387, particularly from a plant which has thegenetic background or architecture at the QTL of line 061M4387, butespecially from a plant of line 061M4387, representative seed of whichis deposited at NCIMB under Accession No. NCIMB 41428, or from a progenyor an ancestor thereof comprising said QTL, and wherein said QTLcontributing to

-   -   a) Bemisia resistance in the donor plant is genetically linked        to at least one marker locus, particularly to at least two        marker loci, more particularly to at least three marker loci and        even more particularly to at least four marker loci, but        especially to at least five and up to six marker loci, which        marker loci are on chromosome 3 and co-segregate with the        Bemisia resistance trait and can be identified by a pair of PCR        oligonucleotide primers selected from the group of primer pairs        1 to 6 represented by a forward and a reverse primer as given in        SEQ ID NOs: 1 to 12; and    -   b) thrips resistance in the donor plant is genetically linked to        at least one marker locus, particularly to at least two marker        loci, particularly to at least three marker loci and        particularly to at least four marker loci, particularly to at        least five marker loci, particularly to at least six marker        loci, and up to seven marker loci, which marker loci are on        chromosome 5 and co-segregate with the thrips resistance trait        and can be identified by a pair of PCR oligonucleotide primers        selected from the group of primer pairs 7 to 13 represented by a        forward and a reverse primer as given in SEQ ID NOs: 13 to 26.

In one embodiment of the invention, one or more primers or probes,particularly one or more primer pairs, but especially one or more primerpairs consisting of a forward primer and a reverse primer, may beestablished for identifying the marker loci according to the inventionby using said one or more primers or probes or said one or more primerpairs, particularly by combining the forward and reverse primers of SEQID NOs: 1-12 to result in a primer pair allowing to identify one or moreof the marker loci on chromosome 3, which co-segregate with the Bemisiaresistance trait.

In one embodiment of the invention, one or more primers or probes,particularly one or more primer pairs, but especially one or more primerpairs consisting of a forward primer and a reverse primer, may beestablished for identifying the marker loci according to the inventionby using said one or more primers or probes or said one or more primerpairs, particularly by combining the forward and reverse primers of SEQID NOs: 13-26 to result in a primer pair allowing to identify one ormore of the marker loci on chromosome 5, which co-segregate with thethrips and/or Bemisia resistance trait.

Also comprised by the present invention are primers and/or probes,particularly primer pairs, but especially primer pairs consisting offorward and reverse primers exhibiting a nucleotide sequence which is atleast 90%, at least 95%, at least 96%, at least 97%, at least 98%, or atleast 99% identical to that given in SEQ ID NOs: 112 and in SEQ ID NOs:13-26, respectively, and also the primer pairs resulting from acombination of said forward and reverse primers.

In particular, the Bemisia resistance trait according to the inventionresiding on chromosome 3 can be identified by a pair of PCRoligonucleotide primers selected from the group of primer pair 1represented by a forward primer which has at least 90%, particularly atleast 95%, particularly at least 96%, particularly at least 97%,particularly at least 98%, particularly at least 99% sequence identitywith the sequence depicted in SEQ ID NO: 1 and a reverse primer whichhas at least 90%, particularly at least 95%, particularly at least 96%,particularly at least 97%, particularly at least 98%, particularly atleast 99% sequence identity with the sequence depicted in SEQ ID NO: 2identifying marker locus 1; primer pair 2 represented by a forwardprimer which has at least 90%, particularly at least 95%, particularlyat least 96%, particularly at least 97%, particularly at least 98%,particularly at least 99% sequence identity with the sequence depictedin SEQ ID NO: 3 and a reverse primer which has at least 90%,particularly at least 95%, particularly at least 96%, particularly atleast 97%, particularly at least 98%, particularly at least 99% sequenceidentity with the sequence depicted in SEQ ID NO: 4 identifying markerlocus 2; primer pair 3 represented by a forward primer which has atleast 90%, particularly at least 95%, particularly at least 96%,particularly at least 97%, particularly at least 98%, particularly atleast 99% sequence identity with the sequence depicted in SEQ ID NO: 5and a reverse primer which has at least 90%, particularly at least 95%,particularly at least 96%, particularly at least 97%, particularly atleast 98%, particularly at least 99% sequence identity with the sequencedepicted in SEQ ID NO: 6 identifying marker locus 3; primer pair 4represented by a forward primer which has at least 90%, particularly atleast 95%, particularly at least 96%, particularly at least 97%,particularly at least 98%, particularly at least 99% sequence identitywith the sequence depicted in SEQ ID NO: 7 and a reverse primer whichhas at least 90%, particularly at least 95%, particularly at least 96%,particularly at least 97%, particularly at least 98%, particularly atleast 99% sequence identity with the sequence depicted in SEQ ID NO: 8identifying marker locus 4; primer pair 5 represented by a forwardprimer which has at least 90%, particularly at least 95%, particularlyat least 96%, particularly at least 97%, particularly at least 98%,particularly at least 99% sequence identity with the sequence depictedin SEQ ID NO: 9 and a reverse primer which has at least 90%,particularly at least 95%, particularly at least 96%, particularly atleast 97%, particularly at least 98%, particularly at least 99% sequenceidentity with the sequence depicted in SEQ ID NO: 10 identifying markerlocus 5; and primer pair 6 represented by a forward primer which has atleast 90%, particularly at least 95%, particularly at least 96%,particularly at least 97%, particularly at least 98%, particularly atleast 99% sequence identity with the sequence depicted in SEQ ID NO: 11and a reverse primer which has at least 90%, particularly at least 95%,particularly at least 96%, particularly at least 97%, particularly atleast 98%, particularly at least 99% sequence identity with the sequencedepicted in SEQ ID NO: 12 identifying marker locus 6, or by any otherprimer or primer pair that identifies a marker locus on chromosome 3that is statistically correlated to the Bemisia resistance trait.

In one embodiment, the thrips resistance trait and/or the Bemisiaresistance trait according to the invention residing on chromosome 5 canbe identified by a pair of PCR oligonucleotide primers selected from thegroup of primer pair 7 represented by a forward primer which has atleast 90%, particularly at least 95%, particularly at least 96%,particularly at least 97%, particularly at least 98%, particularly atleast 99% sequence identity with the sequence depicted in SEQ ID NO: 13and a reverse primer which has at least 90%, particularly at least 95%,particularly at least 96%, particularly at least 97%, particularly atleast 98%, particularly at least 99% sequence identity with the sequencedepicted in SEQ ID NO: 14, identifying marker locus 7; primer pair 8represented by a forward primer which has at least 90%, particularly atleast 95%, particularly at least 96%, particularly at least 97%,particularly at least 98%, particularly at least 99% sequence identitywith the sequence depicted in SEQ ID NO: 15 and a reverse primer whichhas at least 90%, particularly at least 95%, particularly at least 96%,particularly at least 97%, particularly at least 98%, particularly atleast 99% sequence identity with the sequence depicted in SEQ ID NO: 16,identifying marker locus 8; primer pair 9 represented by a forwardprimer which has at least 95%, particularly at least 96%, particularlyat least 97%, particularly at least 98%, particularly at least 99%sequence identity with the sequence depicted in SEQ ID NO: 17 and areverse primer which has at least 95%, particularly at least 96%,particularly at least 97%, particularly at least 98%, particularly atleast 99% sequence identity with the sequence depicted in SEQ ID NO: 18,identifying marker locus 9; primer pair 10 represented by a forwardprimer which has at least 90%, particularly at least 95%, particularlyat least 96%, particularly at least 97%, particularly at least 98%,particularly at least 99% sequence identity with the sequence depictedin SEQ ID NO: 19 and a reverse primer which has at least 90%,particularly at least 95%, particularly at least 96%, particularly atleast 97%, particularly at least 98%, particularly at least 99% sequenceidentity with the sequence depicted in SEQ ID NO: 20, identifying markerlocus 10; primer pair 11 represented by a forward primer which has atleast 90%, particularly at least 95%, particularly at least 96%,particularly at least 97%, particularly at least 98%, particularly atleast 99% sequence identity with the sequence depicted in SEQ ID NO: 21and a reverse primer which has at least 90%, particularly at least 95%,particularly at least 96%, particularly at least 97%, particularly atleast 98%, particularly at least 99% sequence identity with the sequencedepicted in SEQ ID NO: 22, identifying marker locus 11; primer pair 12represented by a forward primer which has at least 90%, particularly atleast 95%, particularly at least 96%, particularly at least 97%,particularly at least 98%, particularly at least 99% sequence identitywith the sequence depicted in SEQ ID NO: 23 and a reverse primer whichhas at least 90%, particularly at least 95%, particularly at least 96%,particularly at least 97%, particularly at least 98%, particularly atleast 99% sequence identity with the sequence depicted in SEQ ID NO: 24,identifying marker locus 12, and primer pair 13 represented by a forwardprimer which has at least 90%, particularly at least 95%, particularlyat least 96%, particularly at least 97%, particularly at least 98%,particularly 99% sequence identity with the sequence depicted in SEQ IDNO: 25 and a reverse primer which has at least 90%, particularly atleast 95%, particularly at least 96%, particularly at least 97%,particularly at least 98%, particularly at least 99% sequence identitywith the sequence depicted in SEQ ID NO: 26, identifying marker locus13; or by any other primer or primer pair that identifies a marker locuson chromosome 5 that is statistically correlated to the thrips and/orBemisia resistance trait.

In one embodiment of the invention oligonucleotide primers are embraced,particularly primer pairs, but especially primer pairs consisting of aforward and a reverse primer exhibiting a nucleotide sequence thathybridizes to the nucleotide sequences of the forward and reverse primersequences given in SEQ ID NOs: 1-12 shown in Table 10 and to thenucleotide sequences of the forward and reverse primer sequences givenin SEQ ID NOs: 13-26 shown in Table 11, respectively, under medium,particularly under medium to high, particularly under high stringencyconditions.

In one embodiment, the invention relates to oligonucleotide sequences,particularly to oligonucleotide sequences that may be used as primersand/or probes, particularly to primer pairs, but especially to primerpairs consisting of a forward and a reverse primer exhibiting anucleotide sequence that hybridizes to nucleotide sequences obtainableby using a forward and a reverse primer exhibiting a nucleotide sequencethat hybridizes to the nucleotide sequences of the forward and reverseprimer sequences given in SEQ ID NOs: 1-12 shown in Table 10 and to thenucleotide sequences of the forward and reverse primer sequences givenin SEQ ID NOs: 13-26 shown in Table 11, respectively, under medium,particularly under medium to high, particularly under high stringencyconditions.

In another embodiment of the invention, a cultivated Capsicum annuumplant is provided as described herein before, wherein said plantcomprises a quantitative trait locus (“QTL”) associated with resistanceto Bemisia, which QTL is characterized by being genetically linked to atleast one marker locus, particularly a marker locus on chromosome 3, andwherein said QTL is further defined by at least one marker allele atsaid at least one marker locus linked to the QTL, which marker allele ischaracterized by the PCR amplification product of an oligonucleotideprimer or primer pair selected from the group of primer pairs 1-6represented by forward and reverse primers of SEQ ID NOs: 1-12,including primer pairs resulting from a combination of the forward andreverse primers of SEQ ID NOs: 1-12, or of any other primer or primerpair that identifies a marker locus on chromosome 3 that isstatistically correlated to the Bemisia resistance trait, whichamplification product corresponds to an amplification product obtainablefrom inbred line 061M4387 (NCIMB 41428) in a PCR reaction with identicalprimers obtainable from said primer pairs 1-6 provided that therespective marker locus is still present in said Capsicum plant.

In particular, the cultivated Capsicum annuum plant as described hereinbefore comprises a quantitative trait locus (“QTL”) associated withresistance to Bemisia, which QTL is characterized by being geneticallylinked to at least one marker locus, particularly a marker locus onchromosome 3, and wherein said QTL is further defined by at least onemarker allele at said at least one marker locus linked to the QTL, whichmarker allele is characterized by the PCR amplification product of anoligonucleotide primer pair selected from the group of primer pair 1represented by a forward primer of SEQ ID NO: 1 and a reverse primer ofSEQ ID NO: 2, identifying marker locus 1; primer pair 2 represented by aforward primer of SEQ ID NO: 3 and a reverse primer of SEQ ID NO: 4,identifying marker locus 2; primer pair 3 represented by a forwardprimer of SEQ ID NO: 5 and a reverse primer of SEQ ID NO: 6, identifyingmarker locus 3; primer pair 4 represented by a forward primer of SEQ IDNO: 7 and a reverse primer of SEQ ID NO: 8, identifying marker locus 4;primer pair 5 represented by a forward primer of SEQ ID NO: 9 and areverse primer of SEQ ID NO: 10, identifying marker locus 5; and primerpair 6 represented by a forward primer of SEQ ID NO: 11 and a reverseprimer of SEQ ID NO: 12, identifying marker locus 6, which amplificationproduct corresponds to an amplification product obtainable from inbredline 061M4387 (NCIMB 41428) in a PCR reaction with primer pairs 1-6identified above provided that the respective marker locus is stillpresent in said Capsicum plant and/or can be considered an allelethereof.

In another embodiment of the invention, a cultivated Capsicum annuumplant is provided as described herein before, wherein said plantcomprises a quantitative trait locus (“QTL”) associated with resistanceto Bemisia, which QTL is characterized by being genetically linked to atleast one marker locus, particularly a marker locus on chromosome 5, andwherein said QTL is further defined by at least one marker allele atsaid at least one marker locus linked to the QTL, which marker allele ischaracterized by the PCR amplification product of an oligonucleotideprimer or primer pair selected from the group of primer pairs 7-13represented by forward and reverse primers of SEQ ID NOs: 13-26,including primer pairs resulting from a combination of the forward andreverse primers of SEQ ID NOs: 13-26, or of any other primer or primerpair that identifies a marker locus on chromosome 5 that isstatistically correlated to the Bemisia resistance trait, whichamplification product corresponds to an amplification product obtainablefrom inbred line 061M4387 (NCIMB 41428) in a PCR reaction with identicalprimers obtainable from primer pairs 7-13 identified above provided thatthe respective marker locus is still present in said Capsicum plantand/or can be considered an allele thereof.

In particular, the cultivated Capsicum annuum plant as described hereinbefore comprises a quantitative trait locus (“QTL”) associated withresistance to Bemisia, which QTL is characterized by being geneticallylinked to at least one marker locus, particularly a marker locus onchromosome 5, and wherein said QTL is further defined by at least onemarker allele at said at least one marker locus linked to the QTL, whichmarker allele is characterized by the PCR amplification product of anoligonucleotide primer pair selected from the group of primer pair 7represented by a forward primer of SEQ ID NO: 13 and a reverse primer ofSEQ ID NO: 14, identifying marker locus 7; primer pair 8 represented bya forward primer of SEQ ID NO: 15 and a reverse primer of SEQ ID NO: 16,identifying marker locus 8; primer pair 9 represented by a forwardprimer of SEQ ID NO: 17 and a reverse primer of SEQ ID NO: 18,identifying marker locus 9; primer pair 10 represented by a forwardprimer of SEQ ID NO: 19 and a reverse primer of SEQ ID NO: 20,identifying marker locus 10; primer pair 11 represented by a forwardprimer of SEQ ID NO: 21 and a reverse primer of SEQ ID NO: 22,identifying marker locus 11; primer pair 12 represented by a forwardprimer of SEQ ID NO: 23 and a reverse primer of SEQ ID NO: 24,identifying marker locus 12, and primer pair 13 represented by a forwardprimer of SEQ ID NO: 25 and a reverse primer of SEQ ID NO: 26,identifying marker locus 13, which amplification product corresponds toan amplification product obtainable from inbred line 061M4387 (NCIMB41428) in a PCR reaction with identical primers obtainable from primerpairs 7-13 identified above provided that the respective marker locus isstill present in said Capsicum plant and/or can be considered an allelethereof.

In another embodiment of the invention, a cultivated Capsicum annuumplant is provided as described herein before, wherein said plantcomprises a quantitative trait locus (“QTL”) associated with resistanceto thrips, which QTL is characterized by being genetically linked to atleast one marker locus, particularly a marker locus on chromosome 5, andwherein said QTL is further defined by at least one marker allele atsaid at least one marker locus linked to the QTL, which marker allele ischaracterized by the PCR amplification product of an oligonucleotideprimer or primer pair selected from the group of primer pairs 7-13represented by forward and reverse primers of SEQ ID NOs: 13-26,including primer pairs resulting from a combination of the forward andreverse primers of SEQ ID NOs: 13-26, or of any other primer or primerpair that identifies a marker locus on chromosome 5 that isstatistically correlated to the thrips resistance trait, whichamplification product corresponds to an amplification product obtainablefrom inbred line 061M4387 (NCIMB 41428) in a PCR reaction with identicalprimers obtainable from primer pairs 7-13 identified above provided thatthe respective marker locus is still present in said Capsicum plantand/or can be considered an allele thereof.

In particular, the cultivated Capsicum annuum plant as described hereinbefore comprises a quantitative trait locus (“QTL”) associated withresistance to thrips, which QTL is characterized by being geneticallylinked to at least one marker locus, particularly a marker locus onchromosome 5, and wherein said QTL is further defined by at least onemarker allele at said at least one marker locus linked to the QTL, whichmarker allele is characterized by the PCR amplification product of anoligonucleotide primer or primer pair selected from the group of primerpair 7 represented by a forward primer of SEQ ID NO: 13 and a reverseprimer of SEQ ID NO: 14, identifying marker locus 7; primer pair 8represented by a forward primer of SEQ ID NO: 15 and a reverse primer ofSEQ ID NO: 16, identifying marker locus 8; primer pair 9 represented bya forward primer of SEQ ID NO: 17 and a reverse primer of SEQ ID NO: 18,identifying marker locus 9; primer pair 10 represented by a forwardprimer of SEQ ID NO: 19 and a reverse primer of SEQ ID NO: 20,identifying marker locus 10; primer pair 11 represented by a forwardprimer of SEQ ID NO: 21 and a reverse primer of SEQ ID NO: 22,identifying marker locus 11; primer pair 12 represented by a forwardprimer of SEQ ID NO: 23 and a reverse primer of SEQ ID NO: 24,identifying marker locus 12, and primer pair 13 represented by a forwardprimer of SEQ ID NO: 25 and a reverse primer of SEQ ID NO: 26,identifying marker locus 13, which amplification product corresponds toan amplification product obtainable from inbred line 061M4387 (NCIMB41428) in a PCR reaction with identical primers obtainable from primerpairs 7-13 identified above provided that the respective marker locus isstill present in said Capsicum plant and/or can be considered an allelethereof.

In one embodiment of the invention, a cultivated Capsicum annuum plantis provided as described herein before, wherein said plant comprises atleast one quantitative trait locus (“QTL”) associated with resistance toBemisia and one quantitative trait locus (“QTL”) associated withresistance to thrips, respectively, which OIL are characterized by beinggenetically linked to at least one marker locus each, particularly amarker locus on chromosome 3 and chromosome 5, respectively, and whereinsaid QTL are further defined by at least one marker allele at said atleast one marker locus linked to

-   -   a) a first QTL on chromosome 3, which marker allele is        characterized by the PCR amplification product of an        oligonucleotide primer or primer pair selected from the group of        primer pairs 1-6 represented by forward and reverse primers of        SEQ ID NOs: 1-12, or of any other primer or primer pair that        identifies a marker locus on chromosome 3 that is statistically        correlated to the Bemisia resistance trait, and    -   b) a second OIL on chromosome 5, which marker allele is        characterized by the PCR amplification product of        oligonucleotide primer or primer pair selected from the group of        primer pairs 7-13 represented by forward and reverse primers of        SEQ ID NOs: 13-26, respectively, or of any other primer or        primer pair that identifies a marker locus on chromosome 5 that        is statistically correlated to the Bemisia and/or thirps        resistance trait,        including primer pairs resulting from a combination of the        forward and reverse primers of SEQ ID NOs: 1-12 and from a        combination of the forward and reverse primers of SEQ ID NOs:        13-26, respectively, which amplification product corresponds to        an amplification product obtainable from inbred line 061M4387        (NCIMB 41428) in a PCR reaction with identical primers        obtainable from said primer pairs 1-6 and 7-13, respectively,        provided that the respective marker locus is still present in        said Capsicum plant and/or can be considered an allele thereof.

In particular, the cultivated Capsicum annuum plant as described hereinbefore comprises a quantitative trait locus (“QTL”) associated

-   -   a) with resistance to Bemisia, which OIL is characterized by        being genetically linked to at least one marker locus,        particularly a marker locus on chromosome 3, and wherein said        OIL is further defined by at least one marker allele at said at        least one marker locus linked to the OIL, which marker allele is        characterized by the PCR amplification product of an        oligonucleotide primer pair selected from the group of primer        pair 1 represented by a forward primer of SEQ ID NO: 1 and a        reverse primer of SEQ ID NO: 2, identifying marker locus 1;        primer pair 2 represented by a forward primer of SEQ ID NO: 3        and a reverse primer of SEQ ID NO: 4, identifying marker locus        2; primer pair 3 represented by a forward primer of SEQ ID NO: 5        and a reverse primer of SEQ ID NO: 6, identifying marker locus        3; primer pair 4 represented by a forward primer of SEQ ID NO: 7        and a reverse primer of SEQ ID NO: 8, identifying marker locus        4; primer pair 5 represented by a forward primer of SEQ ID NO: 9        and a reverse primer of SEQ ID NO: 10, identifying marker locus        5; and primer pair 6 represented by a forward primer of SEQ ID        NO: 11 and a reverse primer of SEQ ID NO: 12, identifying marker        locus 6, or by any other marker locus on chromosome 3 that is        statistically correlated to the Bemisia resistance trait; and    -   b) with resistance to thrips and/or Bemisia, which OIL is        characterized by being genetically linked to at least one marker        locus, particularly a marker locus on chromosome 5, and wherein        said QTL is further defined by at least one marker allele at        said at least one marker locus linked to the QTL, which marker        allele is characterized by the PCR amplification product of an        oligonucleotide primer pair selected from the group of primer        pair 7 represented by a forward primer of SEQ ID NO: 13 and a        reverse primer of SEQ ID NO: 14, identifying marker locus 7;        primer pair 8 represented by a forward primer of SEQ ID NO: 15        and a reverse primer of SEQ ID NO: 16, identifying marker locus        8; primer pair 9 represented by a forward primer of SEQ ID NO:        17 and a reverse primer of SEQ ID NO: 18, identifying marker        locus 9; primer pair 10 represented by a forward primer of SEQ        ID NO: 19 and a reverse primer of SEQ ID NO: 20, identifying        marker locus 10; primer pair 11 represented by a forward primer        of SEQ ID NO: 21 and a reverse primer of SEQ ID NO: 22,        identifying marker locus 11; primer pair 12 represented by a        forward primer of SEQ ID NO: 23 and a reverse primer of SEQ ID        NO: 24, identifying marker locus 12, and primer pair 13        represented by a forward primer of SEQ ID NO: 25 and a reverse        primer of SEQ ID NO: 26, identifying marker locus 13, or by any        other marker locus on chromosome 5 that is statistically        correlated to the thrips and/or Bemisia resistance trait,    -   wherein each amplification product corresponds to an        amplification product obtainable from inbred line 061M4387        (NCIMB 41428) in a PCR reaction with identical primers        obtainable from primer pairs 1-6 and 7-13, respectively,        identified above provided that the respective marker locus is        still present in said Capsicum plant and/or can be considered an        allele thereof.

In one embodiment, the invention relates to the amplification productobtainable in a PCR reaction involving an oligonucleotide primer orprimer pair selected from the group of primer pairs 1-6 represented byforward and reverse primers of SEQ ID NOs: 1-12 and primer pairs 7-13represented by forward and reverse primers of SEQ ID NOs: 13-26,respectively, including primer pairs resulting from a combination of theforward and reverse primers of SEQ ID NOs: 1-12 and SEQ ID NOs: 13-26,respectively, or any other primer or primer pair that identifies amarker locus on chromosome 3 and/or chromosome 5 that is statisticallycorrelated to the Bemisia and/or thrips resistance trait, whichamplification product corresponds to an amplification product obtainablefrom inbred line 061M4387 (NCIMB 41428) in a PCR reaction with identicalprimers obtainable from said primer pairs 1-6 and 7-13, respectively, orcombination of primer pairs provided that the respective marker locus isstill present in said Capsicum plant and/or can be considered an allelethereof.

Also included is a polynucleotide that has at least 90%, particularly atleast 95%, particularly at least 96%, particularly at least 97%,particularly at least 98%, particularly at least 99% sequence identitywith the sequence of said amplification product.

In one embodiment of the invention a polynucleotide is embracedexhibiting a nucleotide sequence that hybridizes to the nucleotidesequences of an amplification product obtainable in a PCR reactioninvolving an oligonucleotide primer pair selected from the group ofprimer pairs 1-6 represented by forward and reverse primers of SEQ IDNOs: 1-12 and primer pairs 7-13 represented by forward and reverseprimers of SEQ ID NOs: 13-26, respectively, including primer pairsresulting from a combination of the forward and reverse primers of SEQID NOs: 1-12 and SEQ ID NOs: 13-26, respectively.

In a specific embodiment, the invention relates to the amplificationproduct obtainable in a PCR reaction involving an oligonucleotide primerpair selected from the group of primer pair 1 represented by a forwardprimer of SEQ ED NO: 1 and a reverse primer of SEQ ID NO: 2, identifyingmarker locus 1; primer pair 2 represented by a forward primer of SEQ IDNO: 3 and a reverse primer of SEQ ID NO: 4, identifying marker locus 2;primer pair 3 represented by a forward primer of SEQ ID NO: 5 and areverse primer of SEQ ID NO: 6, identifying marker locus 3; primer pair4 represented by a forward primer of SEQ ID NO: 7 and a reverse primerof SEQ ID NO: 8, identifying marker locus 4; primer pair 5 representedby a forward primer of SEQ ID NO: 9 and a reverse primer of SEQ ID NO:10, identifying marker locus 5; and primer pair 6 represented by aforward primer of SEQ ID NO: 11 and a reverse primer of SEQ ID NO: 12,identifying marker locus 6, which amplification product corresponds toan amplification product obtainable from inbred line 061M4387 (NCIMB41428) in a PCR reaction with identical primers obtainable from saidprimer pairs 1-6 identified above provided that the respective markerlocus is still present in said Capsicum plant.

In another specific embodiment, the invention relates to theamplification product obtainable in a PCR reaction involving anoligonucleotide primer pair selected from the group of primer pair 7represented by a forward primer of SEQ ID NO: 13 and a reverse primer ofSEQ ID NO: 14, identifying marker locus 7; primer pair 8 represented bya forward primer of SEQ ID NO: 15 and a reverse primer of SEQ ID NO: 16,identifying marker locus 8; primer pair 9 represented by a forwardprimer of SEQ ID NO: 17 and a reverse primer of SEQ ID NO: 18,identifying marker locus 9; primer pair 10 represented by a forwardprimer of SEQ ID NO: 19 and a reverse primer of SEQ ID NO: 20,identifying marker locus 10; primer pair 11 represented by a forwardprimer of SEQ ID NO: 21 and a reverse primer of SEQ ID NO: 22,identifying marker locus 11; primer pair 12 represented by a forwardprimer of SEQ ID NO: 23 and a reverse primer of SEQ ID NO: 24,identifying marker locus 12, and primer pair 13 represented by a forwardprimer of SEQ ID NO: 25 and a reverse primer of SEQ ID NO: 26,identifying marker locus 13, which amplification product corresponds toan amplification product obtainable from inbred line 061M4387 (NCIMB41428) in a PCR reaction with identical primers obtainable from saidprimer pairs 7-13 identified above provided that the respective markerlocus is still present in said Capsicum plant.

The amplification product according to the invention and describedherein above can then be used for generating new primers or probes thatcan be used for identifying a marker locus, particularly a marker locuson chromosome 3 and/or 5, respectively, genetically linked with a QTLassociated with resistance to Bemisia and/or thrips.

In one embodiment the invention relates to a marker, particularly toprimers or probes developed from an amplification product according tothe invention and as described herein above by methods known in the art.

In one embodiment of the invention, a cultivated Capsicum annuum plantaccording to the invention and as described herein before is provided,wherein said allele or alleles associated with resistance to Bemisia isobtainable from line 061M4387, or any other line having the same geneticarchitecture at the QTL on chromosome 3 and/or chromosome 5,representative seed of which is deposited under Accession No. NCIMB41428, or from a progeny or an ancestor thereof comprising said QTL, orQTL architecture.

This would also cover plants where the markers specifically disclosedherein are recombined off and thus no longer present in plant genome.

In one embodiment of the invention, a cultivated Capsicum annuum plantaccording to the invention and as described herein before is provided,wherein said allele associated with resistance to thrips is obtainablefrom line 061M4387, or any other line having the same geneticarchitecture at the QTL on chromosome 5, representative seed of which isdeposited under Accession No. NCIMB 41428, or from a progeny or anancestor thereof comprising said QTL, or QTL architecture.

This would also cover plants where the markers specifically disclosedherein are recombined off and thus no longer present in plant genome.

In one aspect of the invention, the cultivated Capsicum annuum plantaccording to the invention and as described herein before isheterozygous for the Bemisia and/or thrips resistance trait.

In one aspect of the invention, the cultivated Capsicum annuum plantaccording to the invention and as described herein before is homozygousfor the Bemisia and/or thrips resistance trait.

In one aspect of the invention, the cultivated Capsicum annum plantaccording to the invention and as described herein before is homozygousfor a c locus (Blum et al. (2002) Genome, 45: 702-705) or a pun1 allele(Stewart et al. (2005) The Plant Journal, 42: 675-688) or for both a clocus and a pun1 allele.

In still another aspect of the invention, the plant according to theinvention and as described herein before carries fruit, which, atmaturity, weigh over 2 grams or are longer than 1 cm and have a diameterof over 0.5 cm and do not show feeding damage caused by thrips and/orBemisia, when said plant is grown under growing conditions generallyused by growers in regular cropping practice, in open field or ingreenhouse.

The plant according to the invention and as described herein before maybe a sweet pepper plant, a bell pepper, a big rectangular pepper, aconical pepper, a long conical pepper or a blocky-type pepper. The fruitof said plant may be an evergreen, a yellow, orange, ivory or red fruit.

The plant according to the invention may be a hot pepper plant, e.g. amildly pungent pepper used for the fresh market and for processingincluding the long, heart-shaped, thin-fleshed Ancho-type and the long,blunt-ended, thin-fleshed Tuscan-type pepper, the slightly more pungentChili pepper fruit with medium flesh thickness, and a pungent pepperused in both the fresh market and for processing including the long,cylindrical-thick fleshed Jalapeno, the small, slender, tapering Serranoand the irregularly shaped, thin-fleshed Cayenne pepper.

The plant according to the invention and as described herein before maybe an inbred, a dihaploid or a hybrid and/or a male sterile.

In one embodiment, the invention relates to plant material obtainablefrom a plant according to the invention and as described herein beforeincluding, but without being limited thereto, leaves, stems, roots,flowers or flower parts, fruits, pollen, egg cells, zygotes, seeds,cuttings, cell or tissue cultures, or any other part or product of theplant which still exhibits the resistant phenotype according to theinvention, particularly when grown into a plant.

The invention further relates to plant parts obtainable from a plantaccording to the invention and as described herein before including, butwithout being limited thereto, plant seed, plant organs such as, forexample, a root, stem, leaf, flower bud, or embryo, etc, ovules, pollenmicrospores, plant cells, plant tissue, plant cells cultures such as,for example, protoplasts, cell culture cells, cells in plant tissues,pollen, pollen tubes, ovules, embryo sacs, zygotes and embryos atvarious stages of development, etc; which still exhibits the resistantphenotype according to the invention, particularly when grown into aplant.

In one aspect, the invention relates to a method for introducing anallele at a quantitative trait locus (“QTL”) contributing to resistanceto Bemisia into a Capsicum annum plant lacking said allele comprising:

-   -   a) obtaining a first plant of the genus Capsicum according to        the invention and as described herein before, particularly a        Capsicum annuum plant, comprising        -   i) at least one marker allele characterized by the PCR            amplification product, particularly a PCR amplification            product obtainable by using PCR oligonucleotide primers or a            pair of PCR oligonucleotide primers, particularly primer            pairs selected from the group of primer pair 1 represented            by a forward primer of SEQ ID NO: 1 and a reverse primer of            SEQ ID NO: 2, identifying marker locus 1; primer pair 2            represented by a forward primer of SEQ ID NO: 3 and a            reverse primer of SEQ ID NO: 4, identifying marker locus 2;            primer pair 3 represented by a forward primer of SEQ ID NO:            5 and a reverse primer of SEQ ID NO: 6, identifying marker            locus 3; primer pair 4 represented by a forward primer of            SEQ ID NO: 7 and a reverse primer of SEQ ID NO: 8,            identifying marker locus 4; primer pair 5 represented by a            forward primer of SEQ ID NO: 9 and a reverse primer of SEQ            ID NO: 10, identifying marker locus 5; and primer pair 6            represented by a forward primer of SEQ ID NO: 11 and a            reverse primer of SEQ ID NO: 12, identifying marker locus 6;            or any other primer or primer pair that identifies a marker            locus on chromosome 3 that is statistically correlated to            the Bemisia resistance trait; or        -   ii) at least one marker allele characterized by the PCR            amplification product, particularly a PCR amplification            product obtainable by using PCR oligonucleotide primers or a            pair of PCR oligonucleotide primers, particularly primer            pairs selected from the group of primer pair 7 represented            by a forward primer of SEQ ID NO: 13 and a reverse primer of            SEQ ID NO: 14, identifying marker locus 7; primer pair 8            represented by a forward primer of SEQ ID NO: 15 and a            reverse primer of SEQ ID NO: 16, identifying marker locus 8;            primer pair 9 represented by a forward primer of SEQ ID NO:            17 and a reverse primer of SEQ ID NO: 18, identifying marker            locus 9; primer pair 10 represented by a forward primer of            SEQ ID NO: 19 and a reverse primer of SEQ ID NO: 20,            identifying marker locus 10; primer pair 11 represented by a            forward primer of SEQ ID NO: 21 and a reverse primer of SEQ            ID NO: 22, identifying marker locus 11; primer pair 12            represented by a forward primer of SEQ ID NO: 23 and a            reverse primer of SEQ ID NO: 24, identifying marker locus            12, and primer pair 13 represented by a forward primer of            SEQ ID NO: 25 and a reverse primer of SEQ ID NO: 26,            identifying marker locus 13; or any other primer or primer            pair that identifies a marker locus on chromosome 5 that is            statistically correlated to the Bemisia resistance trait or        -   iii) a combination of i) and ii);    -   b) crossing said first plant of the genus Capsicum, particularly        a Capsicum annuum plant, with a second Capsicum annuum plant,        wherein said second Capsicum annum plant lacks said nucleic        acid; and    -   c) identifying a plant resulting from the cross exhibiting        increased resistance to Bemisia and comprising said at least one        marker allele; or, optionally,    -   d) identifying a plant resulting from the cross exhibiting        increased resistance to Bemisia and missing said marker alleles        identified in step a).

In one aspect, the invention relates to a method for introducing anallele at a quantitative trait locus (“QTL”) contributing to resistanceto thrips into a Capsicum annum plant lacking said allele comprising:

-   -   a) obtaining a first plant of the genus Capsicum, particularly a        Capsicum annuum plant, comprising at least one marker allele        characterized by the PCR amplification product, particularly a        PCR amplification product obtainable by using PCR        oligonucleotide primers or a pair of PCR oligonucleotide        primers, particularly primer pairs selected from the group of        primer pair 7 represented by a forward primer of SEQ ID NO: 13        and a reverse primer of SEQ ID NO: 14, identifying marker locus        7; primer pair 8 represented by a forward primer of SEQ ID NO:        15 and a reverse primer of SEQ ID NO: 16, identifying marker        locus 8; primer pair 9 represented by a forward primer of SEQ ID        NO: 17 and a reverse primer of SEQ ID NO: 18, identifying marker        locus 9; primer pair 10 represented by a forward primer of SEQ        ID NO: 19 and a reverse primer of SEQ ID NO: 20, identifying        marker locus 10; primer pair 11 represented by a forward primer        of SEQ ID NO: 21 and a reverse primer of SEQ ID NO: 22,        identifying marker locus 11; primer pair 12 represented by a        forward primer of SEQ ID NO: 23 and a reverse primer of SEQ ID        NO: 24, identifying marker locus 12, and primer pair 13        represented by a forward primer of SEQ ID NO: 25 and a reverse        primer of SEQ ID NO: 26, identifying marker locus 13; or any        other primer or primer pair that identifies a marker locus on        chromosome 5 that is statistically correlated to the thrips        resistance trait;    -   b) crossing said first plant of the genus Capsicum, particularly        a Capsicum annuum plant, with a second Capsicum annuum plant,        wherein said second Capsicum annum plant lacks said nucleic        acid; and    -   c) identifying a plant resulting from the cross exhibiting        increased resistance to thrips and comprising said at least one        marker allele; or, optionally,    -   d) identifying a plant resulting from the cross exhibiting        increased resistance to Bemisia and missing said marker alleles        identified in step a).

In one aspect, the invention relates to a method for introducing atleast a first allele at a quantitative trait locus (“QTL”) contributingto resistance to Bemisia and at least a second allele at a quantitativetrait locus (“QTL”) contributing to resistance to thrips and/or Bemisia,into a Capsicum annuum plant lacking said alleles comprising:

-   -   a) obtaining a first plant of the genus Capsicum, particularly a        Capsicum annuum plant, comprising        -   i) at least one marker allele characterized by the PCR            amplification product, particularly a PCR amplification            product obtainable by using PCR oligonucleotide primers or a            pair of PCR oligonucleotide primers, particularly primer            pairs selected from the group of primer pair 1 represented            by a forward primer of SEQ ID NO: 1 and a reverse primer of            SEQ ID NO: 2, identifying marker locus 1; primer pair 2            represented by a forward primer of SEQ ID NO: 3 and a            reverse primer of SEQ ID NO: 4, identifying marker locus 2;            primer pair 3 represented by a forward primer of SEQ ID NO:            5 and a reverse primer of SEQ ID NO: 6, identifying marker            locus 3; primer pair 4 represented by a forward primer of            SEQ ID NO: 7 and a reverse primer of SEQ ID NO: 8,            identifying marker locus 4; primer pair 5 represented by a            forward primer of SEQ ID NO: 9 and a reverse primer of SEQ            ID NO: 10, identifying marker locus 5; and primer pair 6            represented by a forward primer of SEQ ID NO: 11 and a            reverse primer of SEQ ID NO: 12, identifying marker locus 6;            or any other primer or primer pair that identifies a marker            locus on chromosome 3 that is statistically correlated to            the Bemisia resistance trait; and        -   ii) at least one marker allele characterized by the PCR            amplification product, particularly a PCR amplification            product obtainable by using PCR oligonucleotide primers or a            pair of PCR oligonucleotide primers, particularly primer            pairs selected from the group of primer pair 7 represented            by a forward primer of SEQ ID NO: 13 and a reverse primer of            SEQ ID NO: 14, identifying marker locus 7; primer pair 8            represented by a forward primer of SEQ ID NO: 15 and a            reverse primer of SEQ ID NO: 16, identifying marker locus 8;            primer pair 9 represented by a forward primer of SEQ ID NO:            17 and a reverse primer of SEQ ID NO: 18, identifying marker            locus 9; primer pair 10 represented by a forward primer of            SEQ ID NO: 19 and a reverse primer of SEQ ID NO: 20,            identifying marker locus 10; primer pair 11 represented by a            forward primer of SEQ ID NO: 21 and a reverse primer of SEQ            ID NO: 22, identifying marker locus 11; primer pair 12            represented by a forward primer of SEQ ID NO: 23 and a            reverse primer of SEQ ID NO: 24, identifying marker locus            12, and primer pair 13 represented by a forward primer of            SEQ ID NO: 25 and a reverse primer of SEQ ID NO: 26,            identifying marker locus 13; or by any other primer or            primer pair that identifies a marker locus on chromosome 5            that is statistically correlated to the thrips and/or            Bemisia resistance trait;    -   b) crossing said first plant of the genus Capsicum, particularly        a Capsicum annum plant, with a second Capsicum annuum plant,        wherein said second Capsicum annuum plant lacks said nucleic        acids; and    -   c) identifying a plant resulting from the cross exhibiting        increased resistance to Bemisia and/or thrips, and comprising at        least two marker alleles co-segregating with said resistance or,        optionally,    -   d) identifying a plant resulting from the cross exhibiting        increased resistance to Bemisia and/or thrips and missing said        marker alleles identified in step a).

In one aspect, the invention relates to a method for introducing a QTLcontributing to resistance to Bemisia into a Capsicum annuum plantlacking said allele comprising:

-   -   a) obtaining a first plant of the genus Capsicum, particularly a        Capsicum annuum plant, which plant contains a genome comprising        a quantitative trait locus (“QTL”) which contributes to Bemisia        resistance, wherein said QTL is characterized by being        genetically linked to at least one marker locus, particularly to        at least two marker loci, more particularly to at least three        marker loci and even more particularly to at least four marker        loci, but especially to at least five and up to six marker loci,        which marker loci are on chromosome 3 and co-segregate with the        Bemisia resistance trait and can be identified by using PCR        oligonucleotide primers or a pair of PCR oligonucleotide        primers, particularly primer pairs selected from the group of        primer pair 1 represented by a forward primer of SEQ ID NO: 1        and a reverse primer of SEQ ID NO: 2, identifying marker locus        1; primer pair 2 represented by a forward primer of SEQ ID NO: 3        and a reverse primer of SEQ ID NO: 4, identifying marker locus        2; primer pair 3 represented by a forward primer of SEQ ID NO: 5        and a reverse primer of SEQ ID NO: 6, identifying marker locus        3; primer pair 4 represented by a forward primer of SEQ ID NO: 7        and a reverse primer of SEQ ID NO: 8, identifying marker locus        4; primer pair 5 represented by a forward primer of SEQ ID NO: 9        and a reverse primer of SEQ ID NO: 10, identifying marker locus        5; and primer pair 6 represented by a forward primer of SEQ ID        NO: 11 and a reverse primer of SEQ ID NO: 12, identifying marker        locus 6; or by any other marker locus on chromosome 3 that is        statistically correlated to the Bemisia resistance trait;    -   b) crossing said first plant of the genus Capsicum, particularly        a Capsicum annum plant, with a second Capsicum annuum plant,        wherein said second Capsicum annuum plant lacks said nucleic        acid; and    -   c) identifying a plant resulting from the cross exhibiting        increased resistance to Bemisia and comprising said QTL.

In another aspect, the invention relates to a method for introducing aQTL contributing to thrips resistance into a Capsicum annuum plantlacking said allele comprising:

-   -   a) obtaining a first plant of the genus Capsicum, particularly a        Capsicum annuum plant, which plant contains a genome comprising        a quantitative trait locus (“QTL”) which contributes to thrips        resistance, wherein said QTL is characterized by being        genetically linked to at least one marker locus, particularly to        at least two marker loci, particularly to at least three marker        loci and particularly to at least four marker loci, particularly        to at least five marker loci, particularly to at least six        marker loci, and up to seven marker loci, which marker loci are        on chromosome 5 and co-segregate with the thrips resistance        trait and can be identified by a pair of PCR oligonucleotide        primers selected from the group of primer pair 7 represented by        a forward primer of SEQ ID NO: 13 and a reverse primer of SEQ ID        NO: 14, identifying marker locus 7; primer pair 8 represented by        a forward primer of SEQ ID NO: 15 and a reverse primer of SEQ ID        NO: 16, identifying marker locus 8; primer pair 9 represented by        a forward primer of SEQ ID NO: 17 and a reverse primer of SEQ ID        NO: 18, identifying marker locus 9; primer pair 10 represented        by a forward primer of SEQ ID NO: 19 and a reverse primer of SEQ        ID NO: 20, identifying marker locus 10; primer pair 11        represented by a forward primer of SEQ ID NO: 21 and a reverse        primer of SEQ ID NO: 22, identifying marker locus 11; primer        pair 12 represented by a forward primer of SEQ ID NO: 23 and a        reverse primer of SEQ ID NO: 24, identifying marker locus 12;        and primer pair 13 represented by a forward primer of SEQ ID NO:        25 and a reverse primer of SEQ ID NO: 26, identifying marker        locus 13, or by any other marker locus on chromosome 5 that is        statistically correlated to the thrips resistance trait;    -   b) crossing said first plant of the genus Capsicum, particularly        a Capsicum annum plant, with a second Capsicum annuum plant,        wherein said second Capsicum annuum plant lacks said nucleic        acid; and    -   c) identifying a plant resulting from the cross exhibiting        increased resistance to thrips and comprising said QTL.

In one aspect, the invention relates to a method for introducing QTLcontributing to resistance to Bemisia and to thrips resistance into aCapsicum annuum plant lacking said allele(s) comprising:

-   -   a) obtaining a first plant of the genus Capsicum, particularly a        Capsicum annuum plant, which plant contains a genome comprising        a quantitative trait locus (“QTL”) which contributes to        -   i) Bemisia resistance, wherein said QTL is characterized by            being genetically linked to at least one marker locus,            particularly to at least two marker loci, more particularly            to at least three marker loci and even more particularly to            at least four marker loci, but especially to at least five            and up to six marker loci, which marker loci are on            chromosome 3 and co-segregate with the Bemisia resistance            trait and can be identified by using PCR oligonucleotide            primers or a pair of PCR oligonucleotide primers,            particularly primer pairs selected from the group of primer            pair 1 represented by a forward primer of SEQ ID NO: 1 and a            reverse primer of SEQ ID NO: 2, identifying marker locus 1;            primer pair 2 represented by a forward primer of SEQ ID NO:            3 and a reverse primer of SEQ ID NO: 4, identifying marker            locus 2; primer pair 3 represented by a forward primer of            SEQ ID NO: 5 and a reverse primer of SEQ ID NO: 6,            identifying marker locus 3; primer pair 4 represented by a            forward primer of SEQ ID NO: 7 and a reverse primer of SEQ            ID NO: 8, identifying marker locus 4; primer pair 5            represented by a forward primer of SEQ ID NO: 9 and a            reverse primer of SEQ ID NO: 10, identifying marker locus 5;            and primer pair 6 represented by a forward primer of SEQ ID            NO: 11 and a reverse primer of SEQ ID NO: 12, identifying            marker locus 6, or by any other marker locus on chromosome 3            that is statistically correlated to the Bemisia resistance            trait; and        -   ii) thrips resistance, wherein said QTL is characterized by            being genetically linked to at least one marker locus,            particularly to at least two marker loci, particularly to at            least three marker loci and particularly to at least four            marker loci, particularly to at least five marker loci,            particularly to at least six marker loci, and up to seven            marker loci, which marker loci are on chromosome 5 and            co-segregate with the thrips resistance trait and can be            identified by using PCR oligonucleotide primers or a pair of            PCR oligonucleotide primers, particularly primer pairs            selected from the group of primer pair 7 represented by a            forward primer of SEQ ID NO: 13 and a reverse primer of SEQ            ID NO: 14, identifying marker locus 7; primer pair 8            represented by a forward primer of SEQ ID NO: 15 and a            reverse primer of SEQ ID NO: 16, identifying marker locus 8;            primer pair 9 represented by a forward primer of SEQ ID NO:            17 and a reverse primer of SEQ ID NO: 18, identifying marker            locus 9; primer pair 10 represented by a forward primer of            SEQ ID NO: 19 and a reverse primer of SEQ ID NO: 20,            identifying marker locus 10; primer pair 11 represented by a            forward primer of SEQ ID NO: 21 and a reverse primer of SEQ            ID NO: 22, identifying marker locus 11; primer pair 12            represented by a forward primer of SEQ ID NO: 23 and a            reverse primer of SEQ ID NO: 24, identifying marker locus            12, and primer pair 13 represented by a forward primer of            SEQ ID NO: 25 and a reverse primer of SEQ ID NO: 26,            identifying marker locus 13, or by any other marker locus on            chromosome 5 that is statistically correlated to the thrips            resistance trait;    -   b) crossing said first plant of the genus Capsicum, particularly        a Capsicum annum plant, with a second Capsicum annuum plant,        wherein said second Capsicum annuum plant lacks said nucleic        acid(s); and    -   c) identifying a plant resulting from the cross exhibiting        increased resistance to Bemisia and thrips, and comprising said        QTL.

In a specific embodiment of the invention the first Capsicum annuumplant is a plant which has the genetic background, but particularly thegenetic architecture at the Bemisia and/or the thrips resistance locus,of line 061M4387, representative seed of which is deposited underAccession No. NCIMB 41428, or a progeny or an ancestor thereof,particularly a plant which has the genetic background or architecture atthe QTL of line 061M4387, or a progeny or an ancestor thereof, butespecially a plant of said line 061M4387, or a progeny or an ancestorthereof.

In another specific embodiment of the invention, the identification of aCapsicum plant exhibiting increased resistance to Bemisia and/or thripsand comprising a QTL according to the invention in step c) of any of themethods described herein before is carried out by phenotypic evaluationusing a ranking scale as disclosed in Examples 2A and 2B, respectively,or by using a molecular marker according to the invention and asdisclosed herein before, or a combination thereof.

In one aspect, the invention relates to the use of an allele or a QTLobtainable from a plant which has the genetic background, butparticularly the genetic architecture at the Bemisia and/or the thripsresistance locus, of line 061M4387, representative seed of which isdeposited under Accession No. NCIMB 41428, or a progeny or an ancestorthereof, particularly from a plant which has the genetic architecture atthe QTL contributing to the Bemisia and/or the thrips resistance of line061M4387, or a progeny or an ancestor thereof, but especially from saidline 061M4387, or a progeny or an ancestor thereof, to confer resistanceto Bemisia and/or to thrips upon a Capsicum annum plant lacking saidallele associated with Bemisia resistance and thrips resistance,respectively.

The invention further comprises a cultivated Capsicum annuum plantresistant, particularly intermediately resistant, to Bemisia, whichplant is obtainable by crossing a Capsicum annuum plant susceptible toBemisia with a plant of line 061M4387, representative seed of which isdeposited under Accession No. NCIMB 41428, or a progeny or an ancestorthereof; and by selecting a plant comprising a QTL, which can beidentified in a PCR reaction by a pair of PCR oligonucleotide primersconsisting of a forward primer and a reverse primer exhibiting anucleotide sequence which is at least 90%, particularly at least 95%,particularly at least 96%, particularly at least 97%, particularly atleast 98%, particularly at least 99% identical to that given in SEQ IDNOs: 1-12; or by any other primer or primer pair that identifies amarker locus on chromosome 3 that is statistically correlated to theBemisia resistance trait.

In one embodiment, the invention provides a cultivated Capsicum annuumplant resistant, particularly intermediately resistant, to thrips, whichplant is obtainable by crossing a Capsicum annuum plant susceptible tothrips with a plant of line 061M4387, representative seed of which isdeposited under Accession No. NCIMB 41428, or a progeny or an ancestorthereof; and by selecting a plant comprising a QTL, which can beidentified in a PCR reaction by a pair of PCR oligonucleotide primersconsisting of a forward primer and a reverse primer exhibiting anucleotide sequence which is at least 90%, particularly at least 95%,particularly at least 96%, particularly at least 97%, particularly atleast 98%, particularly at least 99% identical to that given in SEQ IDNOs: 13-26, or by any other primer or primer pair that identifies amarker locus on chromosome 5 that is statistically correlated to thethrips resistance trait.

In one embodiment, the invention provides a cultivated Capsicum annumplant resistant, particularly intermediately resistant, to Bemisia andthrips, which plant is obtainable by crossing a Capsicum annuum plantsusceptible to Bemisia and/or thrips with a plant of line 061M4387,representative seed of which is deposited under Accession No NCIMB41428, or a progeny or an ancestor thereof; and by selecting a plantcomprising a QTL, which can be identified in a PCR reaction by a pair ofPCR oligonucleotide primers consisting of a forward primer and a reverseprimer exhibiting a nucleotide sequence which is at least 90%,particularly at least 95%, particularly at least 96%, particularly atleast 97%, particularly at least 98%, particularly at least 99%identical to that given in SEQ ID NOs: 1-12 and in SEQ ID NOs: 13-26,respectively, or by any other primer or primer pair that identities amarker locus on chromosome 3 and on chromosome 5 that is statisticallycorrelated to the thrips and Bemisia resistance trait, respectively.

In one aspect, the invention relates to a method of producing pepperfruit comprising:

-   -   a) growing a cultivated Capsicum annuum plant resistant,        particularly intermediately resistant, to Bemisia according to        the invention and as described herein before;    -   b) allowing said plant to set fruit; and    -   c) harvesting fruit of said plant.

In one aspect, the invention relates to a method of producing pepperfruit, particularly pepper fruit which is essentially free of feedingdamage caused by Bemisia and/or thrips comprising:

-   -   a) growing a cultivated Capsicum annuum plant resistant,        particularly intermediately resistant, to thrips according to        the invention and as described herein before;    -   b) allowing said plant to set fruit; and    -   c) harvesting fruit of said plant.

In one aspect, the invention relates to a method of producing pepperfruit, particularly pepper fruit which essentially free of feedingdamage caused by thrips comprising:

-   -   a) growing a cultivated Capsicum annuum plant resistant,        particularly intermediately resistant, to Bemisia and to thrips        according to the invention and as described herein before;    -   b) allowing said plant to set fruit; and    -   c) harvesting fruit of said plant.

In another aspect, the invention relates to a method of producing pepperseed comprising:

-   -   a) growing cultivated Capsicum annuum plant resistant,        particularly intermediately resistant, to Bemisia according to        the invention and as described herein before;    -   b) harvesting fruit of said plant; and    -   c) extracting seed from said fruit.

In another aspect, the invention relates to a method of producing pepperseed comprising:

-   -   a) growing cultivated Capsicum annuum plant resistant,        particularly intermediately resistant, to thrips according to        the invention and as described herein before;    -   b) harvesting fruit of said plant; and    -   c) extracting seed from said fruit.

In another aspect, the invention relates to a method of producing pepperseed comprising:

-   -   a) growing cultivated Capsicum annuum plant resistant,        particularly intermediately resistant, to Bemisia and to thrips        according to the invention and as described herein before;    -   b) harvesting fruit of said plant; and    -   c) extracting seed from said fruit.

In one embodiment, the invention relates to a method of producing apepper plant that exhibits resistance to Bemisia, wherein said plantcomprises an allele associated with resistance to Bemisia at aquantitative trait loci (“QTL”) contributing to Bemisia resistancelocated on chromosome 3; particularly at a QTL derived from Capsicumannum line 061M4387 representative seed of which is deposited underAccession No. NCIMB 41428, or from a progeny or an ancestor thereofcomprising said QTL, comprising the steps of:

-   -   a) providing a recipient pepper plant susceptible to Bemisia or        a plant that does not contain a QTL allele conferring resistance        to Bemisia infestations;    -   b) providing a donor pepper plant exhibiting resistance to        Bemisia infestations due to the presence of the resistance QTL        allele on chromosome 3;    -   c) crossing the recipient and the donor plant to produce progeny        plants segregating for the presence of the favourable QTL        allele;    -   d) screening the genome of progeny plants for recombinations in        the region of the QTL on chromosome 3.

In one embodiment, the invention relates to a method of producing apepper plant that exhibits resistance to Bemisia, wherein said plantcomprises an allele associated with resistance to Bemisia at aquantitative trait loci (“QTL”) contributing to thrips resistancelocated on chromosome 3 and/or chromosome 5; particularly at a QTLderived from Capsicum annuum line 061M4387 representative seed of whichis deposited under Accession No, NUMB 41428, or from a progeny or anancestor thereof comprising said QTL, comprising the steps of:

-   -   a) providing a recipient pepper plant susceptible to Bemisia or        a plant that does not contain a QTL allele conferring resistance        to Bemisia infestations;    -   b) providing a donor pepper plant exhibiting resistance to        Bemisia infestations due to the presence of the resistance QTL        allele on chromosome 3 and/or chromosome 5;    -   c) crossing the recipient and the donor plant to produce progeny        plants segregating for the presence of the favourable QTL        allele;    -   d) screening the genome of progeny plants for recombinations in        the region of the QTL on chromosome 3 and/or chromosome 5.

In one embodiment, the invention relates to a method of producing apepper plant that exhibits resistance to thrips, wherein said plantcomprises an allele associated with resistance to thrips at aquantitative trait loci (“QTL”) contributing to thrips resistancelocated on chromosome 5; particularly at a QTL derived from Capsicumannuum line 061M4387 representative seed of which is deposited underAccession No. NUMB 41428, or from a progeny or an ancestor thereofcomprising said QTL, comprising the steps of:

-   -   a) providing a recipient pepper plant susceptible to thrips or a        plant that does not contain a QTL allele conferring resistance        to thrips infestations;    -   b) providing a donor pepper plant exhibiting resistance to        thrips infestations due to the presence of the resistance QTL        allele on chromosome 5;    -   c) crossing the recipient and the donor plant to produce progeny        plants segregating for the presence of the favourable QTL        allele;    -   d) screening the genome of progeny plants for recombinations in        the region of the QTL on chromosome 5.

In one embodiment, the invention relates to a method of producing apepper plant that exhibits resistance to Bemisia, and thrips whereinsaid plant comprises an allele associated with resistance to Bemisia ata quantitative trait loci (“QTL”) contributing to Bemisia resistancelocated on chromosome 3 and an allele associated with resistance tothrips and/or Bemisia at a quantitative trait loci (“QTL”) contributingto thrips and/or Bemisia resistance located on chromosome 5;particularly at QTL located on chromosome 3 and 5, respectively, derivedfrom Capsicum annuum line 061M4387 representative seed of which isdeposited under Accession No. NUMB 41428, or from a progeny or anancestor thereof comprising said QTL, comprising the steps of:

-   -   a) providing a recipient pepper plant susceptible to Bemisia        and/or thrips or a plant that does not contain a QTL allele        conferring resistance to Bemisia and/or thrips infestations;    -   b) providing a donor pepper plant exhibiting resistance to        Bemisia and/or thrips infestations due to the presence of the        resistance OIL allele on chromosome 3 and/or 5, respectively;    -   c) crossing the recipient and the donor plant to produce progeny        plants segregating for the presence of the favourable QTL        allele;    -   d) screening the genome of progeny plants for recombinations in        the region of the QTL on chromosome 3 and/or 5.

In one embodiment, the invention relates to a method of producing apepper plant that exhibits resistance to Bemisia and/or thrips accordingto the invention and as described herein before, wherein said progenyplant is a plant of a segregating population produced byself-pollination of an F1 plant obtained from said cross in step c), orby crossing an F1 plant obtained from said cross with another pepperplant.

In one embodiment the invention relates to a method of producing apepper plant that exhibits resistance to Bemisia and/or thrips accordingto the invention and as described herein before, wherein the segregatingpopulation obtained in step c) may be subjected to a standard resistanceassay such as, for example, a resistance assay as described in Example2A and 2B, respectively.

In one embodiment, the invention relates to a method of producing apepper plant that exhibits resistance to Bemisia and/or thrips accordingto the invention and as described herein before, wherein the screeningof the progeny plants in step d) is be a marker-based screening, whichmay be supported by performing a resistance assay such as, for example,a resistance assay as described in Example 2A and 2B, respectively.

In one embodiment, the invention relates to a method of producing apepper plant that exhibits resistance to Bemisia and/or thrips accordingto the invention and as described herein before, wherein the screeningof the genome is performed using molecular markers genetically linked tochromosome 3 and/or chromosome 5, respectively.

In one embodiment, the invention relates to a method of producing apepper plant that exhibits resistance to Bemisia according to theinvention and as described herein before, wherein the screening of thegenome is performed using molecular markers genetically linked to atleast one marker characterized by the PCR amplification product of a PCRoligonucleotide primer or a pair of PCR oligonucleotide primers selectedfrom the group of primer pairs 1-6 represented by forward and reverseprimers of SEQ ID NOs: 1-12.

In one embodiment, the invention relates to a method of producing apepper plant that exhibits resistance to thrips according to theinvention and as described herein before, wherein the screening of thegenome is performed using molecular markers genetically linked to atleast one marker characterized by the PCR amplification product of a PCRoligonucleotide primer or a pair of PCR oligonucleotide primers selectedfrom the group of primer pairs 7-13 represented by forward and reverseprimers of SEQ ID NOs: 13-26.

In one embodiment, the invention relates to a method of producing apepper plant that exhibits resistance to Bemisia according to theinvention and as described herein before, wherein progeny plants areidentified and selected which comprise a reduced genome segment at theQTL still conferring resistance to Bemisia, wherein the link to at leastone marker allele at least one marker locus linked to the QTL in thedonor plant, which marker allele is characterized by the PCRamplification product of an PCR oligonucleotide primer oroligonucleotide primer pair selected from the group of primer pairs 1-6represented by forward and reverse primers of SEQ ID NOs: 1-12, has beenbroken.

In one embodiment, the invention relates to a method of producing apepper plant that exhibits resistance to Bemisia according to theinvention and as described herein before, wherein progeny plants areidentified and selected which comprise a reduced genome segment at theQTL still conferring resistance to Bemisia, wherein the link to all themarker alleles characterized by the PCR amplification product of an PCRoligonucleotide primer or oligonucleotide primer pair selected from thegroup of primer pairs 1-6 represented by forward and reverse primers ofSEQ ID NOs: 1-12, has been broken.

In one embodiment, the invention relates to a method of producing apepper plant that exhibits resistance to thrips according to theinvention and as described herein before, wherein progeny plants areidentified and selected which comprise a reduced genome segment at theQTL still conferring resistance to thrips, wherein the link to at leastone marker allele at least one marker locus linked to the OIL in thedonor plant, which marker allele is characterized by the PCRamplification product of an PCR oligonucleotide primer oroligonucleotide primer pair selected from the group of primer pairs 7-13represented by forward and reverse primers of SEQ ID NOs: 13-26, hasbeen broken.

In one embodiment, the invention relates to a method of producing apepper plant that exhibits resistance to thrips according to theinvention and as described herein before, wherein progeny plants areidentified and selected which comprise a reduced genome segment at theQTL still conferring resistance to thrips, wherein the link to all themarker alleles characterized by the PCR amplification product of an PCRoligonucleotide primer or oligonucleotide primer pair selected from thegroup of primer pairs 7-13 represented by forward and reverse primers ofSEQ ID NOs: 13-26, has been broken.

In one embodiment, the invention relates to a method of producing apepper plant that exhibits resistance to Bemisia and/or thrips accordingto the invention and as described herein before, wherein plantscomprising a genome segment, particularly a reduced genome segment atthe QTL still conferring resistance to Bemisia and/or thrips areidentified by a marker-based screening or by performing a resistanceassay or by a combination of both, particularly by a marker-basedscreening, wherein markers are used that are located in the QTL regionand are genetically linked to at least one marker locus, provided thatsaid markers are segregating in the same population.

In one embodiment, the invention relates to a method of identifying aquantitative trait locus (“QTL”) which contributes to Bemisia resistancecomprising using in a PCR reaction a PCR oligonucleotide primer or apair of PCR oligonucleotide primers selected from the group of primerpair 1 represented by a forward primer of SEQ ID NO: 1 and a reverseprimer of SEQ ID NO: 2, identifying marker locus 1; primer pair 2represented by a forward primer of SEQ ID NO: 3 and a reverse primer ofSEQ ID NO: 4, identifying marker locus 2; primer pair 3 represented by aforward primer of SEQ ID NO: 5 and a reverse primer of SEQ ID NO: 6,identifying marker locus 3; primer pair 4 represented by a forwardprimer of SEQ ID NO: 7 and a reverse primer of SEQ ID NO: 8, identifyingmarker locus 4; primer pair 5 represented by a forward primer of SEQ IDNO: 9 and a reverse primer of SEQ ID NO: 10, identifying marker locus 5;and primer pair 6 represented by a forward primer of SEQ ID NO: 11 and areverse primer of SEQ ID NO: 12, identifying marker locus 6.

In one embodiment, the invention relates to a method of identifying aquantitative trait locus (“QTL”) which contributes to Bemisia resistancecomprising using in a PCR reaction a PCR oligonucleotide primer or apair of PCR oligonucleotide primers selected from the group of primerpair 7 represented by a forward primer of SEQ ID NO: 13 and a reverseprimer of SEQ ID NO: 14, identifying marker locus 7; primer pair 8represented by a forward primer of SEQ ID NO: 15 and a reverse primer ofSEQ ID NO: 16, identifying marker locus 8; primer pair 9 represented bya forward primer of SEQ ID NO: 17 and a reverse primer of SEQ ID NO: 18,identifying marker locus 9; primer pair 10 represented by a forwardprimer of SEQ ID NO: 19 and a reverse primer of SEQ ID NO: 20,identifying marker locus 10; primer pair 11 represented by a forwardprimer of SEQ ID NO: 21 and a reverse primer of SEQ ID NO: 22,identifying marker locus 11; primer pair 12 represented by a forwardprimer of SEQ ID NO: 23 and a reverse primer of SEQ ID NO: 24,identifying marker locus 12, and primer pair 13 represented by a forwardprimer of SEQ ID NO: 25 and a reverse primer of SEQ ID NO: 26,identifying marker locus 13.

In one embodiment, the invention relates to a method of identifying aquantitative trait locus (“QTL”) which contributes to Bemisia resistancecomprising using in a PCR reaction

-   -   a) a PCR oligonucleotide primer or a pair of PCR oligonucleotide        primers selected from the group of primer pair 1 represented by        a forward primer of SEQ ID NO: 1 and a reverse primer of SEQ ID        NO: 2, identifying marker locus 1; primer pair 2 represented by        a forward primer of SEQ ID NO: 3 and a reverse primer of SEQ ID        NO: 4, identifying marker locus 2; primer pair 3 represented by        a forward primer of SEQ ID NO: 5 and a reverse primer of SEQ ID        NO: 6, identifying marker locus 3; primer pair 4 represented by        a forward primer of SEQ ID NO: 7 and a reverse primer of SEQ ID        NO: 8, identifying marker locus 4; primer pair 5 represented by        a forward primer of SEQ ID NO: 9 and a reverse primer of SEQ ID        NO: 10, identifying marker locus 5; and primer pair 6        represented by a forward primer of SEQ ID NO: 11 and a reverse        primer of SEQ ID NO: 12, identifying marker locus 6; and    -   b) a PCR oligonucleotide primer or a pair of PCR oligonucleotide        primers selected from the group of primer pair 7 represented by        a forward primer of SEQ ID NO: 13 and a reverse primer of SEQ ID        NO: 14, identifying marker locus 7; primer pair 8 represented by        a forward primer of SEQ ID NO: 15 and a reverse primer of SEQ ID        NO: 16, identifying marker locus 8; primer pair 9 represented by        a forward primer of SEQ ID NO: 17 and a reverse primer of SEQ ID        NO: 18, identifying marker locus 9; primer pair 10 represented        by a forward primer of SEQ ID NO: 19 and a reverse primer of SEQ        ID NO: 20, identifying marker locus 10; primer pair 11        represented by a forward primer of SEQ ID NO: 21 and a reverse        primer of SEQ ID NO: 22, identifying marker locus 11; primer        pair 12 represented by a forward primer of SEQ ID NO: 23 and a        reverse primer of SEQ ID NO: 24, identifying marker locus 12,        and primer pair 13 represented by a forward primer of SEQ ID NO:        25 and a reverse primer of SEQ ID NO: 26, identifying marker        locus 13.

In one embodiment, the invention relates to a method of identifying aquantitative trait locus (“QTL”) which contributes to thrips resistancecomprising using in a PCR reaction a PCR oligonucleotide primer or apair of PCR oligonucleotide primers selected from the group of primerpair 7 represented by a forward primer of SEQ ID NO: 13 and a reverseprimer of SEQ ID NO: 14, identifying marker locus 7; primer pair 8represented by a forward primer of SEQ ID NO: 15 and a reverse primer ofSEQ ID NO: 16, identifying marker locus 8; primer pair 9 represented bya forward primer of SEQ ID NO: 17 and a reverse primer of SEQ ID NO: 18,identifying marker locus 9; primer pair 10 represented by a forwardprimer of SEQ ID NO: 19 and a reverse primer of SEQ ID NO: 20,identifying marker locus 10; primer pair 11 represented by a forwardprimer of SEQ ID NO: 21 and a reverse primer of SEQ ID NO: 22,identifying marker locus 11; primer pair 12 represented by a forwardprimer of SEQ ID NO: 23 and a reverse primer of SEQ ID NO: 24,identifying marker locus 12, and primer pair 13 represented by a forwardprimer of SEQ ID NO: 25 and a reverse primer of SEQ ID NO: 26,identifying marker locus 13.

In one embodiment, the invention relates to a QTL identified by a methodaccording to the invention and as described herein before, which QTLcontributes to Bemisia resistance and is located on chromosome 3.

In one embodiment, the invention relates to a QTL identified by a methodaccording to the invention and as described herein before, which QTLcontributes to Bemisia resistance and is located on chromosome 5.

In one embodiment, the invention relates to a QTL identified by a methodaccording to the invention and as described herein before, which QTLcontributes to thrips resistance and is located on chromosome 5.

In one embodiment, the invention provides a cultivated Capsicum annuumplant comprising a genome comprising at least one QTL which contributesto Bemisia resistance, which QTL is located on chromosome 3, whereinsaid at least one QTL can be identified by a molecular marker that is inlinkage disequilibrium and/or linked to and/or located in the QTLregion, as well as a marker that represent the actual causal mutationsunderlying the QTL, and thus exhibits statistical correlation to thephenotypic trait, which marker can be developed using theoligonucleotide primers as disclosed in SEQ ID NO: 1-12.

In one embodiment, the invention provides a cultivated Capsicum annuumplant comprising a genome comprising at least two QTL which contributeto Bemisia resistance, which QTL are located on chromosome 3 and 5,wherein said at least two QTL can be identified by molecular markersthat are in linkage disequilibrium and/or linked to and/or located inthe QTL region, as well as a markers that represent the actual causalmutations underlying the QTL, and thus exhibits statistical correlationto the phenotypic trait, which markers can be developed using theoligonucleotide primers as disclosed in SEQ ID NO: 1-12 and SEQ ID NOs:13 to 26, respectively.

In one embodiment, the invention provides a cultivated Capsicum annuumplant comprising a genome comprising at least one QTL which contributesto thrips resistance, which QTL is located on chromosome 5, wherein saidat least one QTL can be identified by a molecular marker that is inlinkage disequilibrium and/or linked to and/or located in the QTLregion, as well as a marker that represent the actual causal mutationsunderlying the QTL, and thus exhibits statistical correlation to thephenotypic trait, which marker can be developed using theoligonucleotide primers as disclosed in SEQ ID NOs: 13 to 26.

In one embodiment, the invention provides a cultivated Capsicum annuumplant comprising a genome comprising at least two QTL which contributeto Bemisia and thrips resistance, which QTL are located on chromosomes 3and 5 and wherein said at least two QTL can be identified by a molecularmarkers that are in linkage disequilibrium and/or linked to and/orlocated in the QTL region, as well as a markers that represent theactual causal mutations underlying the OIL, and thus exhibitsstatistical correlation to the phenotypic trait, which markers can bedeveloped using the oligonucleotide primers as disclosed in SEQ ID NO:1-12 and SEQ ID NOs: 13 to 26, respectively.

DEFINITIONS

The technical terms and expressions used within the scope of thisapplication are generally to be given the meaning commonly applied tothem in the pertinent art of plant breeding and cultivation if nototherwise indicated herein below.

A “cultivated Capsicum annuum” plant is understood within the scope ofthe invention to refer to a plant that is no longer in the natural statebut has been developed by human care and for human use and/orconsumption.

As used in this specification and the appended claims, the singularforms “a”, “an”, and “the” include plural referents unless the contextclearly dictates otherwise. Thus, for example, reference to “a plant”includes one or more plants, and reference to “a cell” includes mixturesof cells, tissues, and the like.

An “allele” is understood within the scope of the invention to refer toalternative or variant forms of various genetic units identical orassociated with different forms of a gene or of any kind of identifiablegenetic element, which are alternative in inheritance because they aresituated at the same locus in homologous chromosomes. Such alternativeor variant forms may be the result of single nucleotide polymorphisms,insertions, inversions, translocations or deletions, or the consequenceof gene regulation caused by for example, by chemical or structuralmodification, transcription regulation or post-translationalmodification/regulation. In a diploid cell or organism, the two allelesof a given gene or genetic element typically occupy corresponding locion a pair of homologous chromosomes.

An allele associated with a quantitative trait may comprise alternativeor variant forms of various genetic units including those that areidentical or associated with a single gene or multiple genes or theirproducts or even a gene disrupting or controlled by a genetic factorcontributing to the phenotype represented by said QR.

As used herein, the term “marker allele” refers to an alternative orvariant form of a genetic unit as defined herein above, when used as amarker to locate genetic loci containing alleles on a chromosome thatcontribute to variability of phenotypic traits.

As used herein, the term “breeding”, and grammatical variants thereof,refer to any process that generates a progeny individual. Breedings canbe sexual or asexual, or any combination thereof. Exemplary non-limitingtypes of breedings include crossings, selfings, doubled haploidderivative generation, and combinations thereof.

As used herein, the phrase “established breeding population” refers to acollection of potential breeding partners produced by and/or used asparents in a breeding program; e.g., a commercial breeding program. Themembers of the established breeding population are typicallywell-characterized genetically and/or phenotypically. For example,several phenotypic traits of interest might have been evaluated, e.g.,under different environmental conditions, at multiple locations, and/orat different times. Alternatively or in addition, one or more geneticloci associated with expression of the phenotypic traits might have beenidentified and one or more of the members of the breeding populationmight have been genotyped with respect to the one or more genetic locias well as with respect to one or more genetic markers that areassociated with the one or more genetic loci.

As used herein, the phrase “diploid individual” refers to an individualthat has two sets of chromosomes, typically one from each of its twoparents. However, it is understood that in some embodiments a diploidindividual can receive its “maternal” and “paternal” sets of chromosomesfrom the same single organism, such as when a plant is selfed to producea subsequent generation of plants.

“Homozygous” is understood within the scope of the invention to refer tolike alleles at one or more corresponding loci on homologouschromosomes.

“Heterozygous” is understood within the scope of the invention to referto unlike alleles at one or more corresponding loci on homologouschromosomes.

“Backcrossing” is understood within the scope of the invention to referto a process in which a hybrid progeny is repeatedly crossed back to oneof the parents. Different recurrent parents may be used in subsequentbackcrosses.

“Locus” is understood within the scope of the invention to refer to aregion on a chromosome, which comprises a gene or any other geneticelement or factor contributing to a trait.

As used herein, “marker locus” refers to a region on a chromosome, whichcomprises a nucleotide or a polynucleotide sequence that is present inan individual's genome and that is associated with one or more loci ofinterest, which may which comprise a gene or any other genetic elementor factor contributing to a trait. “Marker locus” also refers to aregion on a chromosome, which comprises a polynucleotide sequencecomplementary to a genomic sequence, such as a sequence of a nucleicacid used as probes.

“Genetic linkage” is understood within the scope of the invention torefer to an association of characters in inheritance due to location ofgenes in proximity on the same chromosome, measured by percentrecombination between loci (centi-Morgan, cM).

As used herein, the phrase “quantitative trait” refers to a phenotypictrait that can be described numerically (i.e., quantitated orquantified). A quantitative trait typically exhibits continuousvariation between individuals of a population; that is, differences inthe numerical value of the phenotypic trait are slight and grade intoeach other. Frequently, the frequency distribution in a population of aquantitative phenotypic trait exhibits a bell-shaped curve (i.e.,exhibits a normal distribution between two extremes). In the presentcase the quantitative trait exhibits continuous variation betweenindividuals of a population in terms of resistance to insects of thegenus Bemisia and/or the order Thysanoptera, which resistance is scoredby means of a standardized Insect Resistance Assay using a scale from1-9 for assessing the severity of the infestation. A quantitative traitis typically the result of a genetic locus interacting with theenvironment or of multiple genetic loci (QTL) interacting with eachother and/or with the environment. Examples of quantitative traitsinclude plant height and yield.

For the purpose of the present invention, the term “co-segregation”refers to the fact that the allele for the trait and the allele(s) forthe marker(s) tend to be transmitted together because they arephysically close together on the same chromosome (reduced recombinationbetween them because of their physical proximity) resulting in anon-random association of their alleles as a result of their proximityon the same chromosome. “Co-segregation” also refers to the presence oftwo or more traits within a single plant of which at least one is knownto be genetic and which cannot be readily explained by chance.

As used herein, the terms “quantitative trait locus” (QTL) and “markertrait association” refer to an association between a genetic marker anda chromosomal region and/or gene that affects the phenotype of a traitof interest. Typically, this is determined statistically; e.g., based onone or more methods published in the literature. A QTL can be achromosomal region and/or a genetic locus with at least two alleles thatdifferentially affect a phenotypic trait (either a quantitative trait ora qualitative trait).

As used herein, the term “genetic architecture at the QTL.” refers to agenomic region which is statistically correlated to the phenotypic traitof interest and represents the underlying genetic basis of thephenotypic trait of interest.

As used herein, the phrases “sexually crossed” and “sexual reproduction”in the context of the presently disclosed subject matter refers to thefusion of gametes to produce progeny (e.g., by fertilization, such as toproduce seed by pollination in plants). A “sexual cross” or“cross-fertilization” is in some embodiments fertilization of oneindividual by another (e.g., cross-pollination in plants). The term“seifing” refers in some embodiments to the production of seed byself-fertilization or self-pollination; i.e., pollen and ovule are fromthe same plant.

As used herein, the phrase “genetic marker” refers to a feature of anindividual's genome (e.g., a nucleotide or a polynucleotide sequencethat is present in an genome) that is associated with one or more lociof interest. In some embodiments, a genetic marker is polymorphic in apopulation of interest, or the locus occupied by the polymorphism,depending on context. Genetic markers include, for example, singlenucleotide polymorphisms (SNPs), indels (i.e., insertions/deletions),simple sequence repeats (SSRs), restriction fragment lengthpolymorphisms (RFLPs), random amplified polymorphic DNAs (RAPDs),cleaved amplified polymorphic sequence (CAPS) markers, Diversity ArraysTechnology (DArT) markers, and amplified fragment length polymorphisms(AFLPs), among many other examples. Genetic markers can, for example, beused to locate genetic loci containing alleles on a chromosome thatcontribute to variability of phenotypic traits. The phrase “geneticmarker” can also refer to a polynucleotide sequence complementary to agenomic sequence, such as a sequence of a nucleic acid used as probes.

A genetic marker can be physically located in a position on a chromosomethat is within or outside of to the genetic locus with which it isassociated (i.e., is intragenic or extragenic, respectively). Statedanother way, whereas genetic markers are typically employed when thelocation on a chromosome of the gene or of a functional mutation, e.g.within a control element outside of a gene, that corresponds to thelocus of interest has not been identified and there is a non-zero rateof recombination between the genetic marker and the locus of interest,the presently disclosed subject matter can also employ genetic markersthat are physically within the boundaries of a genetic locus (e.g.,inside a genomic sequence that corresponds to a gene such as, but notlimited to a polymorphism within an intron or an exon of a gene). Insome embodiments of the presently disclosed subject matter, the one ormore genetic markers comprise between one and ten markers, and in someembodiments the one or more genetic markers comprise more than tengenetic markers.

As used herein, the term “genotype” refers to the genetic constitutionof a cell or organism. An individual's “genotype for a set of geneticmarkers” includes the specific alleles, for one or more genetic markerloci, present in the individual's haplotype. As is known in the art, agenotype can relate to a single locus or to multiple loci, whether theloci are related or unrelated and/or are linked or unlinked. In someembodiments, an individual's genotype relates to one or more genes thatare related in that the one or more of the genes are involved in theexpression of a phenotype of interest (e.g., a quantitative trait asdefined herein). Thus, in some embodiments a genotype comprises asummary of one or more alleles present within an individual at one ormore genetic loci of a quantitative trait. In some embodiments, agenotype is expressed in terms of a haplotype (defined herein below).

As used herein, the term “germplasm” refers to the totality of thegenotypes of a population or other group of individuals (e.g., aspecies). The term “germplasm” can also refer to plant material; e.g., agroup of plants that act as a repository for various alleles. The phrase“adapted germplasm” refers to plant materials of proven geneticsuperiority; e.g., for a given environment or geographical area, whilethe phrases “non adapted germplasm,” “raw germplasm,” and “exoticgermplasm” refer to plant materials of unknown or unproven geneticvalue; e.g., for a given environment or geographical area; as such, thephrase “non-adapted germplasm” refers in some embodiments to plantmaterials that are not part of an established breeding population andthat do not have a known relationship to a member of the establishedbreeding population.

As used herein, the terms “hybrid”, “hybrid plant,” and “hybrid progeny”refers to an individual produced from genetically different parents(e.g., a genetically heterozygous or mostly heterozygous individual).

As used herein, the phrase “single cross F₁ hybrid” refers to an F₁hybrid produced from a cross between two inbred lines.

As used herein, the phrase “inbred line” refers to a geneticallyhomozygous or nearly homozygous population. An inbred line, for example,can be derived through several cycles of brother/sister breedings or ofsetting or in dihaploid production. In some embodiments, inbred linesbreed true for one or more phenotypic traits of interest. An “inbred”,“Inbred individual”, or “inbred progeny” is an individual sampled froman inbred line.

As used herein, the term “dihaploid line”, refers to stable inbred linesissued from anther culture. Some pollen grains (haploid) cultivated onspecific medium and circumstances can develop plantlets containing nchromosomes. These plantlets are then “doubled” and contain 2nchromosomes. The progeny of these plantlets are named “dihaploid” andare essentially not segregating any more (stable).

As used herein, the term “linkage”, and grammatical variants thereof,refers to the tendency of alleles at different loci on the samechromosome to segregate together more often than would be expected bychance if their transmission were independent, in some embodiments as aconsequence of their physical proximity.

As used herein, the term “locus” refers to a position on a chromosome(e.g., of a gene, a genetic marker, or the like).

As used herein, the phrase “nucleic acid” refers to any physical stringof monomer units that can be corresponded to a string of nucleotides,including a polymer of nucleotides (e.g., a typical DNA, cDNA or RNApolymer), modified oligonucleotides (e.g., oligonucleotides comprisingbases that are not typical to biological RNA or DNA, such as2′-O-methylated oligonucleotides), and the like. In some embodiments, anucleic acid can be single-stranded, double-stranded, multi-stranded, orcombinations thereof. Unless otherwise indicated, a particular nucleicacid sequence of the presently disclosed subject matter optionallycomprises or encodes complementary sequences, in addition to anysequence explicitly indicated.

As used herein, the phrase “phenotypic trait” refers to the appearanceor other detectable characteristic of an individual, resulting from theinteraction of its genome, proteome and/or metabolome with theenvironment.

As used herein, the phrase “resistance” refers to the ability of a plantto restrict the growth and development of a specified pest or pathogenand/or the damage they cause when compared to susceptible plants undersimilar environmental conditions and pest or pathogen pressure.Resistant plants may exhibit some disease symptoms or damage under heavypest or pathogen pressure.

Essentially two levels of resistance are to be distinguished. “High orstandard resistance” refers to plants that highly restrict the growthand development of the specified pest or pathogen under normal pest orpathogen pressure when compared to susceptible counterparts. Theseplants may, however, exhibit some symptoms or damage under heavy pest orpathogen pressure.

“Moderate/intermediate resistance” refers to plants that distractinsects and/or restrict the growth and development of the specified pestor pathogen, or show reduced damage compared to susceptible counterpartsbut may exhibit a greater range of symptoms or damage compared tohigh/standard resistant plants. Moderately/intermediately resistantplants will still show significantly less severe symptoms or damage thansusceptible plants when grown under similar environmental conditionsand/or pest or pathogen pressure.

As used herein, the phrase “susceptibility” refers to the inability of aplant to adequately restrict the growth and development of a specifiedpest or pathogen.

As used herein, the phrase “Bemisia resistance” or “resistance toBemisia infestations” or “Bemisia resistant plant” refers to the plantscapability to resist attack, infestation, or colonization by the insect.The level of resistance exhibited by a certain plant can be scored, forexample, by means of a standardized Insect Resistance Assay as describedin Example 2A herein below using a scale from 1-9 for assessing theseverity of the infestation.

Plants scoring 1 in said Insect Resistance Assay are completely coveredwith pupae and heavily moulded often stunted in growth, whereas plantsscoring 9 are completely free of pupae and thus fully resistant. Astandard “susceptible variety” (e.g. Vergasa F1 or Biking F1) isunderstood for the purpose of the present invention to refer to a plantthat scores in an Insect Resistance Assay as described in Example 2Abetween 3 and 4 where the plants show many pupae (100-400/leaf) whichare densely crowded on the leaf, usually accompanied by black mould.

A “Bemisia resistant plant” is understood for the purpose of the presentinvention to refer to a plant that scores in a standardized InsectResistance Assay as described in Example 2A herein below in a range ofbetween 6 and 9, including 6 and 9.

A moderate or intermediate resistance to Bemisia infestations starts ata score of 6 where the plants show a moderate-relatively low number ofpupae (20-50/leaf) which are more regular distributed over the leaves.At a score of 7 only some pupae (5-20/leaf) are present, which areirregularly scattered over the leaf. Plants scoring 8 show only very few(1-5/leaf) pupae and are not noticeably affected in growth or fruitdevelopment.

Accordingly, for the purpose of the present invention, by a plant being“moderately or intermediately resistant” to Bemisia infestation, a plantis to be understood that scores in the range of between 6 and 8 on ascale ranging from 1-9 determined in a standardized Insect ResistanceAssay as described in Example 2A herein below. A plant is understood tobe “highly resistant” to Bemisia, if it scores in the range of between 8and 9, including 9.

As used herein, the phrase “thrips resistance” or “resistance to thripsinfestations” or “thrips resistant plant” refers to the plantscapability to resist attack, infestation, or colonization by the insect.The level of resistance exhibited by a certain plant can be scored, forexample, by means of a standardized Insect Resistance Assay as describedin Example 2B herein below using a scale from 1-9 for assessing theseverity of the infestation judged on the basis of the observed feedingdamage (silvering).

Plants scoring 1 in said Insect Resistance Assay show very heavysilvering with a large part of the leaf damaged (>40% silvering),whereas plants scoring 9 show no silvering damage (0% silvering) and arethus fully resistant. A standard “susceptible variety” (e.g. Roxy F1and/or Snooker F1) is understood for the purpose of the presentinvention to refer to a plant that scores in an Insect Resistance Assayas described in Example 2B between 3 (11%-20% silvering) and 4 (6%-10%silvering) where the plants show many large silvering spots distributedover the entire leaf.

A “thrips resistant plant” is understood for the purpose of the presentinvention to refer to a plant that scores in a standardized InsectResistance Assay as described in Example 2B herein below in a range ofbetween 5 and 9, including 5 and 9.

A moderate or intermediate resistance to thrips infestations starts at ascore of 5 where the plants show a moderate number of spots more regulardistributed over the leaves (3%-5% silvering). At a score of 7 theplants show only some small spots especially near the mid vein or leafedge (0.1%-1% silvering). Plants scoring 8 show only tiny spots and arenot noticeably affected in growth or fruit development (<0.1%silvering).

Accordingly, for the purpose of the present invention, by a plant being“moderately or intermediately resistant” to thrips infestation, a plantis to be understood that scores in the range of between 5 and 8,particularly between 6 and 8, on a scale ranging from 1-9 determined ina standardized Insect Resistance Assay as described in Example 28 hereinbelow. A plant is understood to be “highly resistant” to thrips, if itscores in the range of between 8 and 9, including 9.

The terms “chromosome 3” and “chromosome 5” are meant to include, andthus used herein synonymously with, the terms “linkage group 3 and 5”and/or “chromosome equivalent of linkage group 3 and 5”, respectively.

As used herein, the term “plurality” refers to more than one. Thus, a“plurality of individuals” refers to at least two individuals. In someembodiments, the term plurality refers to more than half of the whole.For example, in some embodiments a “plurality of a population” refers tomore than half the members of that population.

As used herein, the term “progeny” refers to the descendant(s) of aparticular cross. Typically, progeny result from breeding of twoindividuals, although some species (particularly some plants andhermaphroditic animals) can be selfed (i.e., the same plant acts as thedonor of both male and female gametes). The descendant(s) can be, forexample, of the F₁, the F₂, or any subsequent generation.

As used herein, the phrase “qualitative trait” refers to a phenotypictrait that is controlled by one or a few genes that exhibit majorphenotypic effects. Because of this, qualitative traits are typicallysimply inherited. Examples in plants include, but are not limited to,flower color, fruit color, and several known disease resistances suchas, for example, Bacterial spot resistance.

“Marker-based selection” is understood within the scope of the inventionto refer to e.g. the use of genetic markers to detect one or morenucleic acids from the plant, where the nucleic acid is associated witha desired trait to identify plants that carry genes for desirable (orundesirable) traits, so that those plants can be used (or avoided) in aselective breeding program.

“Microsatellite or SSRs (Simple sequence repeats) Marker” is understoodwithin the scope of the invention to refer to a type of genetic markerthat consists of numerous repeats of short sequences of DNA bases, whichare found at loci throughout the plant's genome and have a likelihood ofbeing highly polymorphic.

“PCR (Polymerase chain reaction)” is understood within the scope of theinvention to refer to a method of producing relatively large amounts ofspecific regions of DNA or subset(s) of the genome, thereby makingpossible various analyses that are based on those regions.

“PCR primer” is understood within the scope of the invention to refer torelatively short fragments of single-stranded DNA used in the PCRamplification of specific regions of DNA.

“Phenotype” is understood within the scope of the invention to refer toa distinguishable characteristic(s) of a genetically controlled trait.

“Polymorphism” is understood within the scope of the invention to referto the presence in a population of two or more different forms of agene, genetic marker, or inherited trait or a gene product obtainable,for example, through alternative splicing, DNA methylation, etc.

“Selective breeding” is understood within the scope of the invention torefer to a program of breeding that uses plants that possess or displaydesirable traits as parents.

“Tester” plant is understood within the scope of the invention to referto a plant of the genus Capsicum used to characterize genetically atrait in a plant to be tested.

Typically, the plant to be tested is crossed with a “tester” plant andthe segregation ratio of the trait in the progeny of the cross isscored.

“Probe” as used herein refers to a group of atoms or molecules which iscapable of recognising and binding to a specific target molecule orcellular structure and thus allowing detection of the target molecule orstructure. Particularly, “probe” refers to a labeled DNA or RNA sequencewhich can be used to detect the presence of and to quantitate acomplementary sequence by molecular hybridization.

The term “hybridize” as used herein refers to conventional hybridizationconditions, preferably to hybridization conditions at which 5×SSPE, 1%SOS, 1×Denhardts solution is used as a solution and/or hybridizationtemperatures are between 35° C. and 70° C., preferably 65° C. Afterhybridization, washing is preferably carried out first with 2×SSC, 1%SOS and subsequently with 0.2×SSC at temperatures between 35° C. and 75°C., particularly between 45° C. and 65° C., but especially at 59° C.(regarding the definition of SSPE, SSC and Denhardts solution seeSambrook et al. loc. cit.). High stringency hybridization conditions asfor instance described in Sambrook et al, supra, are particularlypreferred. Particularly preferred stringent hybridization conditions arefor instance present if hybridization and washing occur at 65° C. asindicated above. Non-stringent hybridization conditions for instancewith hybridization and washing carried out at 45° C. are less preferredand at 35° C. even less.

“Sequence Homology or Sequence Identity” is used herein interchangeably.The terms “identical” or percent “identity” in the context of two ormore nucleic acid or protein sequences, refer to two or more sequencesor subsequences that are the same or have a specified percentage ofamino acid residues or nucleotides that are the same, when compared andaligned for maximum correspondence, as measured using one of thefollowing sequence comparison algorithms or by visual inspection. If twosequences which are to be compared with each other differ in length,sequence identity preferably relates to the percentage of the nucleotideresidues of the shorter sequence which are identical with the nucleotideresidues of the longer sequence. Sequence identity can be determinedconventionally with the use of computer programs such as the Besffitprogram (Wisconsin Sequence Analysis Package, Version 8 for Unix,Genetics Computer Group, University Research Park, 575 Science DriveMadison, Wis. 53711). Bestfit utilizes the local homology algorithm ofSmith and Waterman, Advances in Applied Mathematics 2 (1981), 482-489,in order to find the segment having the highest sequence identitybetween two sequences. When using Bestfit or another sequence alignmentprogram to determine whether a particular sequence has for instance 95%identity with a reference sequence of the present invention, theparameters are preferably so adjusted that the percentage of identity iscalculated over the entire length of the reference sequence and thathomology gaps of up to 5% of the total number of the nucleotides in thereference sequence are permitted. When using Bestfit, the so-calledoptional parameters are preferably left at their preset (“default”)values. The deviations appearing in the comparison between a givensequence and the above-described sequences of the invention may becaused for instance by addition, deletion, substitution, insertion orrecombination. Such a sequence comparison can preferably also be carriedout with the program “fasta20u66” (version 2.0u66, September 1998 byWilliam R. Pearson and the University of Virginia; see also W. R.Pearson (1990), Methods in Enzymology 183, 63-98, appended examples andhttp://workbench.scisc.edu/). For this purpose, the “default” parametersettings may be used.

Another indication that two nucleic acid sequences are substantiallyidentical is that the two molecules hybridize to each other understringent conditions. The phrase: “hybridizing specifically to” refersto the binding, duplexing, or hybridizing of a molecule only to aparticular nucleotide sequence under stringent conditions when thatsequence is present in a complex mixture (e.g., total cellular) DNA orRNA. “Bind(s) substantially” refers to complementary hybridizationbetween a probe nucleic acid and a target nucleic acid and embracesminor mismatches that can be accommodated by reducing the stringency ofthe hybridization media to achieve the desired detection of the targetnucleic acid sequence.

“Stringent hybridization conditions” and “stringent hybridization washconditions” in the context of nucleic acid hybridization experimentssuch as Southern and Northern hybridizations are sequence dependent, andare different under different environmental parameters. Longer sequenceshybridize specifically at higher temperatures. An extensive guide to thehybridization of nucleic acids is found in Tijssen (1993) LaboratoryTechniques in Biochemistry and Molecular Biology-Hybridization withNucleic Acid Probes part I chapter 2 “Overview of principles ofhybridization and the strategy of nucleic acid probe assays” Elsevier,N.Y. Generally, highly stringent hybridization and wash conditions areselected to be about 5° C. lower than the thermal melting point(T.sub.m) for the specific sequence at a defined ionic strength and pH.Typically, under “stringent conditions” a probe will hybridize to itstarget subsequence, but to no other sequences.

The T.sub.m is the temperature (under defined ionic strength and pH) atwhich 50% of the target sequence hybridizes to a perfectly matchedprobe. Very stringent conditions are selected to be equal to the T.sub.mfor a particular probe. An example of stringent hybridization conditionsfor hybridization of complementary nucleic acids which have more than100 complementary residues on a filter in a Southern or northern blot is50% formamide with 1 mg of heparin at 42° C., with the hybridizationbeing carried out overnight. An example of highly stringent washconditions is 0.1 5M NaCl at 72° C. for about 15 minutes. An example ofstringent wash conditions is a 0.2 times SSC wash at 65° C. for 15minutes (see, Sambrook, infra, for a description of SSC buffer). Often,a high stringency wash is preceded by a low stringency wash to removebackground probe signal. An example medium stringency wash for a duplexof, e.g., more than 100 nucleotides, is 1 times SSC at 45° C. for 15minutes. An example low stringency wash for a duplex of, e.g., more than100 nucleotides, is 4-5 times SSC at 40° C. for 15 minutes. For shortprobes (e.g., about 10 to 50 nucleotides), stringent conditionstypically involve salt concentrations of less than about 1.0M Na ion,typically about 0.01 to 1.0 M Na ion concentration (or other salts) atpH 7.0 to 8.3, and the temperature is typically at least about 30° C.Stringent conditions can also be achieved with the addition ofdestabilizing agents such as formamide. In general, a signal to noiseratio of 2 times (or higher) than that observed for an unrelated probein the particular hybridization assay indicates detection of a specifichybridization. Nucleic acids that do not hybridize to each other understringent conditions are still substantially identical if the proteinsthat they encode are substantially identical. This occurs, e.g. when acopy of a nucleic acid is created using the maximum codon degeneracypermitted by the genetic code.

A “plant” is any plant at any stage of development, particularly a seedplant.

A “plant cell” is a structural and physiological unit of a plant,comprising a protoplast and a cell wall. The plant cell may be in formof an isolated single cell or a cultured cell, or as a part of higherorganized unit such as, for example, plant tissue, a plant organ, or awhole plant.

“Plant cell culture” means cultures of plant units such as, for example,protoplasts, cell culture cells, cells in plant tissues, pollen, pollentubes, ovules, embryo sacs, zygotes and embryos at various stages ofdevelopment.

“Plant material” refers to leaves, stems, roots, flowers or flowerparts, fruits, pollen, egg cells, zygotes, seeds, cuttings, cell ortissue cultures, or any other part or product of a plant.

A “plant organ” is a distinct and visibly structured and differentiatedpart of a plant such as a root, stem, leaf, flower bud, or embryo.

“Plant tissue” as used herein means a group of plant cells organizedinto a structural and functional unit. Any tissue of a plant in plantaor in culture is included. This term includes, but is not limited to,whole plants, plant organs, plant seeds, tissue culture and any groupsof plant cells organized into structural and/or functional units. Theuse of this term in conjunction with, or in the absence of, any specifictype of plant tissue as listed above or otherwise embraced by thisdefinition is not intended to be exclusive of any other type of planttissue.

The present invention relates to novel pepper plants, particular toCapsicum annuum plants, resistant, particularly intermediatelyresistant, to insects, particularly to insects of the genus Bemisiaand/or the order Thysanoptera, more particularly to Bemisia tabaci(white fly) and/or trips more particularly Frankliniella occidentalis,further to seeds and fruits of said plants. The present invention alsorelates to methods of making and using such plants and their fruits.

Plants according to the invention may be obtained by crossing two ormore parental genotypes, at least one of which may have one or morealleles, particularly one or more alleles at corresponding QTLcontributing to Bemisia and/or thrips resistance, which allele(s) is/arelacking in the other parental genotype or which complements the othergenotype to obtain a plant according to the invention and as describedherein before. If more than one QTL contributes to the expression of theresistance trait and the two original parental genotypes do not providethe entire set of alleles, other sources can be included in the breedingpopulation. The other parental genotype may contribute a desirable traitincluding fruit quality demanded by the market such as, for example, aweight in the range of 180 grams, blocky shape, smooth skin, bright redcolour. Beside fruit quality, agronomically important characteristicssuch as, for example, a good plant architecture, high productivity andbasic resistances to disease such as, but not limited to, TMV (TobaccoMosaic virus) and TSWV (Tomato Spotted Wilt virus) are further desiredtraits.

These parental genotypes may be crossed with one another to produceprogeny seed. The parental genotypes may be inbred lines developed byselfing selected heterozygous plants from fields with uncontrolled oropen pollination and employing recurrent selection procedures. Superiorplants are selfed and selected in successive generations. In thesucceeding generations the heterozygous condition gives way tohomogeneous lines as a result of self-pollination and selection. Withsuccessive generations of inbreeding, the plant becomes more and morehomozygous and uniform within the progeny plants. Typically, five toseven or more generations (F1 to F2; F3 to F4; F4 to F5) of selfing andpedigree selection may be practiced to obtain inbred lines that areuniform in plant and seed characteristics and that will remain uniformunder continued self-fertilization.

During inbreeding, many undesirable alleles at heterozygous loci will bereplaced by more favourable alleles and the unfavourable or undesiredalleles eliminated from the progeny Moreover, through marker-basedselection the number of favorable alleles can be maximized in that themore unfavourable alleles are identified and successively replaced bythe more favorable alleles.

In one aspect, the plant according to the invention may be obtained byintrogressing the insect resistance trait from an ancestor plant,particularly a wild ancestor plant into a cultivated pepper plant,particularly a cultivated Capsicum annuum plant, more particularly aCapsicum annuum plant which is homozygous for a c locus (Blum et al.(2002) Genome, 45: 702-705) or a pun1 allele (Stewart et al. (2005) ThePlant Journal, 42: 675-688), or for both loci.

In one specific embodiment of the invention, the wild ancestor, fromwhich the Bemisia and/or thrips resistance trait may be obtained, iswild Capsicum annuum accession no CGN16975 obtainable from the Instituutvoor de Veredeling van Tuinbouwgewassen (now: Centre for GeneticResources), Wageningen, Netherlands. The insect resistance traitaccording to the present invention, which confers to a plant expressingthis trait, an intermediate level of resistance to infestations withinsects of the genus Bemisia and/or of the order Thysanoptera, moreparticularly to Bemisia tabaci (white fly) and/or thrips moreparticularly Frankliniella occidentalis, may, in the alternative, beobtained from Capsicum annuum line 061M4387, a sample of which has beendeposited with NCIMB Ltd under accession number NCIMB 41428, or from aprogeny or ancestor of line 061M4387 comprising the Bemisia and/or thethrips resistance trait.

Accordingly, in a specific embodiment of the invention, the parentalgenotype contributing the resistance trait(s) is an inbred line havingthe invention relevant properties of deposited Capsicum annuum line061M4387, i. e. substantially the same genome architecture at the QTLassociated with Bemisia and/or thrips resistance, seed samples of whichhave been deposited on Aug. 10, 2006 with NCIMB under accession numberNCIMB 41428.

In another specific embodiment of the invention, the parental genotypecontributing to the resistance trait is a hybrid having the inventionrelevant properties of deposited Capsicum annuum line 061M4387, i.e.substantially the same genome architecture at the QTL associated withBemisia and/or thrips resistance, seed samples of which have beendeposited on Aug. 10, 2006 with NCIMB under accession number NCIMB41428.

Capsicum annuum line 061M4387 resulted from a cross of wild accession noCGN16975 obtainable from the Centre for Genetic Resources, Wageningen,Netherlands as the donor of the resistance trait with a Capsicum annuuminbred line. Bemisia and thrips resistant progeny of this cross wascrossed with further inbred lines of different genetic backgrounds tofinally obtain line 061M4387.

Accordingly, Capsicum annuum line 061M4387 or any other plant linecontaining the Bemisia and/or thrips resistance trait of Capsicum annuumline 061M4387, may be used as a source material for introgressing saidresistance trait into any desired genetic background to obtain a pepperplant being highly or intermediately resistant, particularlyintermediately resistant, to infestations with insects of the genusBemisia and/or of the order Thysanoptera, more particularly to Bemisiatabaci (white fly) and/or thrips more particularly Frankliniellaoccidentalis, may further contain one or more desirable traits such asfruit quality traits demanded by the market such as, for example, aweight in the range of 180 grams, blocky shape, smooth skin, bright redcolour. Beside fruit quality, agronomically important characteristicssuch as, for example, a good plant architecture, high productivity andbasic resistances to disease such as, but not limited to, TMV (TobaccoMosaic virus) and TSWV (Tomato Spotted Wilt virus) are further desiredtraits.

Based on the description of the present invention, the skilled personwho is in possession of Capsicum annuum line 061M4387, a sample of whichhas been deposited with NCIMB Ltd under accession number NCIMB 41428, orof a progeny or ancestor thereof containing the QTL on chromosome 3associated with resistance to Bemisia and/or the QTL on chromosome 5associated with resistance to thrips and Bemisia, respectively, asdescribed herein above, has no difficulty to transfer the Bemisiaresistance trait and/or the thrips resistance trait of the presentinvention to other pepper plants of various types using breedingtechniques well-known in the art. The trait of the present invention mayfor example be transferred to pepper plants producing fruit of varioustypes or shapes, such as bell peppers, sweet peppers, hot peppers, bigrectangular peppers, conical peppers, including long conical peppers, orblocky-type peppers and of various mature colors, such as evergreen,red, yellow, orange or ivory. Accordingly, in one embodiment, a plant ofthe present invention is a C. annum plant capable of resistinginfestations with Bemisia and/or thrips, which plant is a bell pepper orsweet pepper, a hot pepper, a big rectangular pepper, a conical pepperor a long conical pepper according to the instant invention. In oneembodiment, a plant of the present invention is capable of producing anevergreen, a red, yellow, orange or ivory pepper fruit. In anotherembodiment of the invention, the pepper plants are grown for (hybrid)seed or commercial pepper production.

Accordingly, in another embodiment, the present invention discloses amethod of transferring the Bemisia resistance trait and/or the thripsresistance trait according to the present invention to a pepper plantlacking said trait comprising a) obtaining a plant comprising saidtrait; b) crossing it to a plant lacking said trait; c) obtaining plantsof the cross of step b); d) selecting a plant of step c) which iscapable of resisting infestations with Bemisia and/or thrips accordingto the present invention. In one embodiment, the method furthercomprises e) back-crossing a plant resulting from step d) with a pepperplant, and f) selecting for a pepper plant, which is capable ofresisting infestations with Bemisia and/or thrips according to thepresent invention. In one embodiment, the method further comprisesobtaining an inbred pepper plant, which is capable of resistinginfestations with Bemisia and/or thrips according to the presentinvention, and, in one embodiment, the method further comprises crossingsaid inbred pepper plant to another pepper plant to produce a hybridpepper plant, which is capable of resisting infestations with Bemisiaand/or thrips according to the present invention. In one embodiment, apepper plant is selected by determining the resistance score to Bemisiaand/or thrips infestations, as described herein. In one embodiment, theplant of step a) comprising said trait is Capsicum annuum line 061M4387,a sample of which has been deposited with NCIMB Ltd under accessionnumber NCIMB 41428, or a progeny or ancestor of said plant.

In certain embodiments of the invention, a standardized InsectResistance Assay is used, such as that described in Example 2 hereinbelow, to determine the resistance level of the progeny plants resultingfrom one of the above crosses and to select those progeny plants forfurther breeding which are intermediately resistant, to Bemisia and/orthrips infestations.

In one embodiment, the present invention discloses a C. annuum plantobtainable by any one of the methods above, wherein the plant is capableof resisting infestations with Bemisia and/or thrips as describedherein.

In yet another embodiment, the present invention discloses a method ofproducing a plant comprising the Bemisia and/or the thrips resistancetrait according to the present invention to a pepper plant lacking saidtrait comprising a) obtaining a plant comprising said trait, b) crossingit to a plant lacking said trait; c) obtaining plants of the cross ofstep b), d) selecting a plant of step c) which is capable of resistinginfestations with Bemisia and/or thrips according to the presentinvention. In one embodiment, the method further comprises e)back-crossing a plant resulting from step d) with a pepper plant, and f)selecting for a pepper plant, which is capable of resisting infestationswith Bemisia and/or thrips according to the present invention. In oneembodiment, the method further comprises obtaining an inbred pepperplant, which is capable of infestations with Bemisia and/or thripsaccording to the present invention, and, in one embodiment, the methodfurther comprises crossing said inbred pepper plant to another pepperplant to produce a hybrid pepper plant, which is capable of resistinginfestations with Bemisia and/or thrips according to the presentinvention. In one embodiment, a pepper plant is selected by determiningthe resistance score to Bemisia and/or thrips infestations, as describedherein. In one embodiment, the, the plant of step a) comprising saidtrait is a Capsicum annuum line 061M4387, a sample of which has beendeposited with NCIMB Ltd under accession number NCIMB 41428, or aprogeny or ancestor of said plant.

In one embodiment, the present invention discloses a C. annuum plantobtainable by any one of the methods above, wherein the plant is capableof resisting infestations with Bemisia and/or thrips as describedherein.

Based on the teachings of the present invention, a skilled person candesign a program to look for new sources for a trait, particularly aresistance trait, but especially a resistance to insects of the genusBemisia and/or of the order Thysanoptera.

In one aspect of the invention, plants expressing the insect resistanttrait and exhibiting resistance, particularly an intermediate level ofresistance, to infections with insects of the genus Bemisia, may beidentified and selected by using a standardized Bemisia resistant testresulting in a resistance rating which is commonly used and recognizedin the art of pepper breeding.

In particular, plants are raised and cultivated according to standardprocedures and transplanted according to a special design.

Plants are transplanted in several rows with a fixed number of plantsper row. In each row one side is used as spreader row and planted with asusceptible entry or, susceptible parental line. The other part of therow is planted with the entries to be tested for insect resistance. Thetest entries are fully randomized in each of several blocks, with 1 ormore plants/entry per block.

Bemisia development is monitored in the spreader row weekly or biweeklyby assessing a fixed number of spreader row plants on equidistantpositions (for example, plant 1, 38, 75, 112, 150) in each row. Thefinal assessments are made when the average infestation of the monitoredspreader plants has reached a resistance rating of approx. 4, that iswhen the pupae are densely crowded on the leaf in numbers of more than100/leaf. Usually, this stage is reached at the time when the firstfruits are ripening (3-4 months after transplantation).

Data are analyzed by calculating the means per test entry and comparisonwith a susceptible entry (e.g. susceptible spreader row or else). Amultiple comparison of the means (e.g. LSD) indicates if test entriesdiffer mutually and from a susceptible control.

For assessing the severity, a scale from 1-9 is used (Table 1). Theabaxial side of the leaves of the plant is inspected and the average ofthe ca. 5 worst affected leaves is assessed according the 1-9 scale. Alltest plants are scored in this way.

In one aspect of the invention, plants expressing the insect resistanttrait and exhibiting resistance, particularly an intermediate level ofresistance, to infections with insects of the order Thysanoptera may beidentified and selected by using a standardized thrips resistant testresulting in a resistance rating which is commonly used and recognizedin the art of pepper breeding.

In particular, plants are raised and cultivated according to standardprocedures and transplanted according to a special design.

In the alternative, marker-assisted breeding may be employed to identifythose individuals where invention relevant loci, particularly inventionrelevant QTL loci, and/or flanking marker loci or marker locigenetically linked thereto, as described herein before have favorablegenotypes, particularly homozygous favorable genotypes.

In one embodiment of the invention, resistance to Bemisia and/or thripsinfestation is recorded in phenotypic evaluation.

In another embodiment, selection is based on molecular markers, whichare linked to traits of interest.

In one embodiment, selection is based on a combination of molecularmarkers and phenotypic evaluation.

Marker-based selection may already be used in the early phases of inbreddevelopment, often in combination with screening methods which are basedlargely on phenotypic characteristics that can be determined visuallyand are related to key performance indices such as, for example, plantvigor, length of internodes, ramifications, insect resistance such asresistance to Bemisia and/or thrips infestations, virus resistances suchas TMV (Tobacco Mosaic virus) and TSWV (Tomato Spotted wilt virus),etc., which are relevant for the suitability of the plant to be utilizedin commercial hybrid production. Selection may also be based onmolecular markers, which may or may not be linked to traits of interest.

In particular, marker-based selection may be applied in combination withor followed by a phenotypic selection to identify those individualswhere all of the invention relevant loci described herein before havehomozygous favorable genotypes.

There are several types of molecular markers that may be used inmarker-based selection including, but not limited to, restrictionfragment length polymorphism (RFLP), random amplification of polymorphicDNA (RAPD), amplified restriction fragment length polymorphism (AFLP),single sequence repeats (SSR) and single nucleotide polymorphisms SNPs.

RFLP involves the use of restriction enzymes to cut chromosomal DNA atspecific short restriction sites, polymorphisms result from duplicationsor deletions between the sites or mutations at the restriction sites.

RAPD utilizes low stringency polymerase chain reaction (PCR)amplification with single primers of arbitrary sequence to generatestrain-specific arrays of anonymous DNA fragments. The method requiresonly tiny DNA samples and analyses a large number of polymorphic loci.

AFLP requires digestion of cellular DNA with a restriction enzyme(s)before using PCR and selective nucleotides in the primers to amplifyspecific fragments. With this method, using electrophoresis techniquesto visualize the obtained fragments, up to 100 polymorphic loci can bemeasured per primer combination and only small DNA sample are requiredfor each test.

SSR analysis is based on DNA micro-satellites (short-repeat) sequencesthat are widely dispersed throughout the genome of eukaryotes, which areselectively amplified to detect variations in simple sequence repeats.Only tiny DNA samples are required for an SSR analysis. SNPs use PCRextension assays that efficiently pick up point mutations. The procedurerequires little DNA per sample. One or two of the above methods may beused in a typical marker-based selection breeding program.

The most preferred method of achieving amplification of nucleotidefragments that span a polymorphic region of the plant genome employs thepolymerase chain reaction (“PCR”) (Mullis et al., Cold Spring HarborSymp. Quant. Biol. 51:263 273 (1986)), using primer pairs involving aforward primer and a backward primer that are capable of hybridizing tothe proximal sequences that define a polymorphism in its double-strandedform.

Alternative methods may be employed to amplify fragments, such as the“Ligase Chain Reaction” (“LOR”) (Barony, Proc. Natl. Acad. Sci. (U.S.A.)88:189 193 (1991)), which uses two pairs of oligonucleotide probes toexponentially amplify a specific target. The sequences of each pair ofoligonucleotides are selected to permit the pair to hybridize toabutting sequences of the same strand of the target. Such hybridizationforms a substrate for a template-dependent ligase. As with PCR, theresulting products thus serve as a template in subsequent cycles and anexponential amplification of the desired sequence is obtained.

LCR can be performed with oligonucleotides having the proximal anddistal sequences of the same strand of a polymorphic site. In oneembodiment, either oligonucleotide will be designed to include theactual polymorphic site of the polymorphism. In such an embodiment, thereaction conditions are selected such that the oligonucleotides can beligated together only if the target molecule either contains or lacksthe specific nucleotide that is complementary to the polymorphic sitepresent on the oligonucleotide. Alternatively, the oligonucleotides maybe selected such that they do not include the polymorphic site (see,Segev, POT Application WO 90/01069).

A further method that may alternatively be employed is the“Oligonucleotide Ligation Assay” (“OLA”) (Landegren et al., Science241:1077 1080 (1988)). The OLA protocol uses two oligonucleotides thatare designed to be capable of hybridizing to abutting sequences of asingle strand of a target. OLA, like LCR, is particularly suited for thedetection of point mutations. Unlike LCR, however, OLA results in“linear” rather than exponential amplification of the target sequence.

Still another method that may alternatively be employed is the “InvaderAssay” that uses a structure-specific flap endonuclease (FEN) to cleavea three-dimensional complex formed by hybridization of allele-specificoverlapping oligonucleotides to target DNA containing a singlenucleotide polymorphism (SNP) site Annealing of the oligonucleotidecomplementary to the SNP allele in the target molecule triggers thecleavage of the oligonucleotide by cleavase, a thermostable FEN.Cleavage can be detected by several different approaches. Most commonly,the cleavage product triggers a secondary cleavage reaction on afluorescence resonance energy transfer (FRET) cassette to release afluorescent signal. Alternatively, the cleavage can be detected directlyby use of fluorescence polarization (FP) probes, or by massspectrometry. The invasive cleavage reaction is highly specific, has alow failure rate, and can detect zeptomol quantities of target DNA.While the assay traditionally has been used to interrogate one SNP inone sample per reaction, novel chip- or bead-based approaches have beentested to make this efficient and accurate assay adaptable tomultiplexing and high-throughput SNP genotyping.

Nickerson et al. have described a nucleic acid detection assay thatcombines attributes of PCR and OLA (Nickerson et al., Proc. Natl. Acad.Sci. (U.S.A.) 87:8923 8927 (1990)). In this method, PCR is used toachieve the exponential amplification of target DNA, which is thendetected using OLA.

Schemes based on ligation of two (or more) oligonucleotides in thepresence of a nucleic acid having the sequence of the resulting“di-oligonucleotide”, thereby amplifying the di-oligonucleotide, arealso known (Wu et al., Genomics 4:560 569 (1989)), and may be readilyadapted to the purposes of the present invention.

In one embodiment, a molecular marker is a DNA fragment amplified byPCR, e.g. a SSR marker or a RAPD marker. In one embodiment, the presenceor absence of an amplified DNA fragment is indicative of the presence orabsence of the trait itself or of a particular allele of the trait. Inone embodiment, a difference in the length of an amplified DNA fragmentis indicative of the presence of a particular allele of a trait, andthus enables to distinguish between different alleles of a trait.

In a specific embodiment of the invention simple sequence repeat (SSR)markers are used to identify invention-relevant alleles in the parentplants and/or the ancestors thereof, as well as in the progeny plantsresulting from a cross of said parent plants. Simple sequence repeatsare short, repeated DNA sequences and present in the genomes of alleukaryotes and consists of several to over a hundred repeats of a givennucleotide motif. Since the number of repeats present at a particularlocation in the genome often differs among plants, SSRs can be analyzedto determine the absence or presence of specific alleles.

In another embodiment of the invention SNP markers are used to identifyinvention-relevant alleles in the parent plants and/or the ancestorsthereof, as well as in the progeny plants resulting from a cross of saidparent plants.

In one aspect, the invention relates to a marker or a set of two or moremarkers and up to 6 markers comprising a pair of PCR oligonucleotideprimers consisting of a forward primer and a reverse primer selectedfrom the group of primer pair 1 represented by a forward primer of SEQID NO: 1 and a reverse primer of SEQ ID NO: 2, primer pair 2 representedby a forward primer of SEQ ID NO: 3 and a reverse primer of SEQ ID NO:4, primer pair 3 represented by a forward primer of SEQ ID NO: 5 and areverse primer of SEQ ID NO: 6, primer pair 4 represented by a forwardprimer of SEQ ID NO: 7 and a reverse primer of SEQ ID NO: 8, primer pair5 represented by a forward primer of SEQ ID NO: 9 and a reverse primerof SEQ ID NO: 10, and primer pair 6 represented by a forward primer ofSEQ ID NO: 11 and a reverse primer of SEQ ID NO: 12, which primers leadto an amplification product in a PCR reaction exhibiting a molecularweight or a nucleotide sequence, which is essentially identical or canbe considered as an allele to that of a corresponding PCR amplificationproduct obtainable from Capsicum annuum line 061M4387 in a PCR reactionwith the identical primer pair(s).

Any other combination of forward and reverse primers selected from thegroup of primer sequences depicted in SEQ ID NOs: 1-12 may also be usedin a PCR reaction.

In one aspect, the invention relates to a marker or a set of two or moremarkers and up to 7 markers comprising a pair of PCR oligonucleotideprimers consisting of a forward primer and a reverse primer selectedfrom the group of primer pair 7 represented by a forward primer of SEQID NO: 13 and a reverse primer of SEQ ID NO: 14, identifying markerlocus 7; primer pair 8 represented by a forward primer of SEQ ID NO: 15and a reverse primer of SEQ ID NO: 16, identifying marker locus 8;primer pair 9 represented by a forward primer of SEQ ID NO: 17 and areverse primer of SEQ ID NO: 18, identifying marker locus 9; primer pair10 represented by a forward primer of SEQ ID NO: 19 and a reverse primerof SEQ ID NO: 20, identifying marker locus 10; primer pair 11represented by a forward primer of SEQ ID NO: 21 and a reverse primer ofSEQ ID NO: 22, identifying marker locus 11; primer pair 12 representedby a forward primer of SEQ ID NO: 23 and a reverse primer of SEQ ID NO:24, identifying marker locus 12, and primer pair 13 represented by aforward primer of SEQ ID NO: 25 and a reverse primer of SEQ ID NO: 26,identifying marker locus 13, which primers lead to an amplificationproduct in a PCR reaction exhibiting a molecular weight or a nucleotidesequence, which is essentially identical or can be considered as anallele to that of a corresponding PCR amplification product obtainablefrom Capsicum annuum fine 061M4387 in a PCR reaction with the identicalprimer pair(s).

Any other combination of forward and reverse primers selected from thegroup of primer sequences depicted in SEQ ID NOs: 13-26 may also be usedin a PCR reaction.

In one aspect, the invention relates to a marker or as set of two ormore markers and up to 13 markers comprising a pair of PCRoligonucleotide primers consisting of a forward and a reverse primerselected from the group of primer pair 1 represented by a forward primerof SEQ ID NO: 1 and a reverse primer of SEQ ID NO: 2, primer pair 2represented by a forward primer of SEQ ID NO: 3 and a reverse primer ofSEQ ID NO: 4, primer pair 3 represented by a forward primer of SEQ IDNO: 5 and a reverse primer of SEQ ID NO: 6, primer pair 4 represented bya forward primer of SEQ ID NO: 7 and a reverse primer of SEQ ID NO: 8,primer pair 5 represented by a forward primer of SEQ ID NO: 9 and areverse primer of SEQ ID NO: 10, and primer pair 6 represented by aforward primer of SEQ ID NO: 11 and a reverse primer of SEQ ID NO: 12,primer pair 7 represented by a forward primer of SEQ ID NO: 13 and areverse primer of SEQ ID NO: 14, identifying marker locus 7; primer pair8 represented by a forward primer of SEQ ID NO: 15 and a reverse primerof SEQ ID NO: 16, identifying marker locus 8; primer pair 9 representedby a forward primer of SEQ ID NO: 17 and a reverse primer of SEQ ID NO:18, identifying marker locus 9; primer pair 10 represented by a forwardprimer of SEQ ID NO: 19 and a reverse primer of SEQ ID NO: 20,identifying marker locus 10; primer pair 11 represented by a forwardprimer of SEQ ID NO: 21 and a reverse primer of SEQ ID NO: 22,identifying marker locus 11; primer pair 12 represented by a forwardprimer of SEQ ID NO: 23 and a reverse primer of SEQ ID NO: 24,identifying marker locus 12, and primer pair 13 represented by a forwardprimer of SEQ ID NO: 25 and a reverse primer of SEQ ID NO: 26,identifying marker locus 13, which primers lead to an amplificationproduct in a PCR reaction exhibiting a molecular weight or a nucleotidesequence, which is essentially identical or can be considered as anallele to that of a corresponding PCR amplification product obtainablefrom Capsicum annuum line 061M4387 in a PCR reaction with the identicalprimer pair(s).

Any other combination of forward and reverse primers selected from thegroup of primer sequences depicted in SEQ ID NOs: 1-26 may also be usedin a PCR reaction.

In another embodiment of the invention, molecular markers may be usedthat are in linkage disequilibrium and/or linked to and/or located inthe QTL region on chromosome 3 and chromosome 5, respectively comprisinga QTL contributing to Bemisia and thirps resistance according to theinvention, as well as a markers that represent the actual causalmutations underlying the QTL, and thus exhibits statistical correlationto the phenotypic trait, which markers can be developed using theoligonucleotide primers as disclosed in SEQ ID NO: 1-12 and SEQ ID NOs:13 to 26, respectively.

In a first step, DNA or cDNA samples are obtained from suitable plantmaterial such as leaf tissue by extracting DNA or RNA using knowntechniques. Primers that flank a region containing SSRs within theinvention-relevant QTL disclosed herein before or within a region linkedthereto, are then used to amplify the DNA sample using the polymerasechain reaction (PCR) method well-known to those skilled in the art.

Basically, the method of PCR amplification involves use of a primer or apair of primers comprising two short oligonucleotide primer sequencesflanking the DNA segment to be amplified or adapter sequences ligated tosaid DNA segment. Repeated cycles of heating and denaturation of the DNAare followed by annealing of the primers to their complementarysequences at low temperatures, and extension of the annealed primerswith DNA polymerase. The primers hybridize to opposite strands of theDNA target sequences. Hybridization refers to annealing of complementaryDNA strands, where complementary refers to the sequence of thenucleotides such that the nucleotides of one strand can bond with thenucleotides on the opposite strand to form double stranded structures.The primers are oriented so that DNA synthesis by the polymeraseproceeds bidirectionally across the nucleotide sequence between theprimers. This procedure effectively doubles the amount of that DNAsegment in one cycle. Because the PCR products are complementary to, andcapable of binding to, the primers, each successive cycle doubles theamount of DNA synthesized in the previous cycle. The result of thisprocedure is exponential accumulation of a specific target fragment,that is approximately 2<n>, where n is the number of cycles.

Through PCR amplification millions of copies of the DNA segment flankedby the primers are made. Differences in the number of repeated sequencesor insertions or deletions in the region flanking said repeats, whichare located between the flanking primers in different alleles arereflected in length variations of the amplified DNA fragments. Thesevariations can be detected, for example, by electrophoreticallyseparating the amplified DNA fragments on gels or by using capillarysequencer. By analyzing the gel or profile, it can be determined whetherthe plant contains the desired allele in a homozygous or heterozygousstate or whether the desired or undesired allele is absent from theplant genome.

Marker analysis can be done early in plant development using DNA samplesextracted from leaf tissue of very young plants or from seed. Thisallows to identify plants with a desirable genetic make-up early in thebreeding cycle and to discard plants that do not contain the desired,invention-relevant alleles prior to pollination thus reducing the sizeof the breeding population and reducing the requirements of phenotyping.

Further, by using molecular markers, a distinction can be made betweenhomozygous plants that carry two copies of the desired,invention-relevant allele at invention-relevant QTL loci andheterozygous plants that carry only one copy and plants that do notcontain any copy of the favourable allele(s).

In one embodiment of the invention, the marker loci can be identified bya pair of PCR oligonucleotide primers consisting of a forward primer anda reverse primer selected from the group of primer pair 1 represented bya forward primer of SEQ ID NO: 1 and a reverse primer of SEQ ID NO: 2,primer pair 2 represented by a forward primer of SEQ ID NO: 3 and areverse primer of SEQ ID NO: 4, primer pair 3 represented by a forwardprimer of SEQ ID NO: 5 and a reverse primer of SEQ ID NO: 6, primer pair4 represented by a forward primer of SEQ ID NO: 7 and a reverse primerof SEQ ID NO: 8, primer pair 5 represented by a forward primer of SEQ IDNO: 9 and a reverse primer of SEQ ID NO: 10, and primer pair 6represented by a forward primer of SEQ ID NO: 11 and a reverse primer ofSEQ ID NO: 12, including oligonucleotide primers consisting of a forwardprimer and a reverse primer exhibiting a nucleotide sequences that sharebetween 90% and 99%, particularly between 95% and 98% sequence identitywith the nucleotide sequences given in SEQ ID NO: 1-12.

In one embodiment of the invention, the marker loci can be identified bya pair of PCR oligonucleotide primers consisting of a forward primer anda reverse primer selected from the group of primer pair 7 represented bya forward primer of SEQ ID NO: 13 and a reverse primer of SEQ ID NO: 14;identifying marker locus 7; primer pair 8 represented by a forwardprimer of SEQ ID NO: 15 and a reverse primer of SEQ ID NO: 16,identifying marker locus 8; primer pair 9 represented by a forwardprimer of SEQ ID NO: 17 and a reverse primer of SEQ ID NO: 18,identifying marker locus 9; primer pair 10 represented by a forwardprimer of SEQ ID NO: 19 and a reverse primer of SEQ ID NO: 20,identifying marker locus 10; primer pair 11 represented by a forwardprimer of SEQ ID NO: 21 and a reverse primer of SEQ ID NO: 22,identifying marker locus 11; primer pair 12 represented by a forwardprimer of SEQ ID NO: 23 and a reverse primer of SEQ ID NO: 24,identifying marker locus 12, and primer pair 13 represented by a forwardprimer of SEQ ID NO: 25 and a reverse primer of SEQ ID NO: 26,identifying marker locus 13, including oligonucleotide primersconsisting of a forward primer and a reverse primer exhibiting anucleotide sequences that share between 90% and 99%, particularlybetween 95% and 98% sequence identity with the nucleotide sequencesgiven in SEQ ID NO: 13-26.

Further can be used within the scope of the invention oligonucleotidemolecules such as primers or probes, particularly primers consisting ofa forward primer and a reverse primer exhibiting a nucleotide sequencesthat hybridize to the nucleotide sequences of the forward and reverseprimer sequences given in SEQ ID NO: 1-12 shown in Table 10 and in SEQID NO: 13-26 shown in Table 11, or to nucleotide sequences thathybridize to a sequence which can be obtained by using forward andreverse primer sequences as given in SEQ ID NO: 1-12 and SEQ ID NO:13-26, respectively, under medium, particularly under medium to high,particularly under high stringency conditions.

In particular, the hybridization reaction is carried out under highstringency conditions at which 5×SSPE, 1% SDS, 1×Denhardts solution isused as a solution and/or hybridization temperatures are between 35° C.and 70° C., and up to 72° C., preferably 65° C. After hybridization,washing is particularly carried out first with 2×SSC, 1% SDS andsubsequently with 0.2×SSC at temperatures between 35° C. and 70° C., andup to 72° C., particularly at 65° C. (regarding the definition of SSPE,SSC and Denhardts solution see Sambrook et al. loc. cit.).

In one aspect of the invention, markers maybe developed and used whichare not explicitly disclosed herein or markers even yet to beidentified. Based on the information provided in this application itwill be possible, for a skilled person, to identify or develop markersnot explicitly disclosed but linked to the QTL or linked to the markersdisclosed. The skilled person knows that other markers may provide atleast equal utility in marker assisted selection.

The invention thus also relates to molecular markers that are in linkagedisequilibrium and/or linked to and/or located in the QTL region onchromosome 3 and chromosome 5, respectively comprising a QTLcontributing to Bemisia and thirps resistance according to theinvention, as well as a markers that represent the actual causalmutations underlying the QTL, and thus exhibits statistical correlationto the phenotypic trait, which markers can be developed using theoligonucleotide primers as disclosed in in SEQ ID NO: 1-12 and SEQ IDNOs: 13 to 26, respectively.

The contiguous genomic markers that indicate the location of the QTL onthe genome are in principal arbitrary or non-limiting. In general, thelocation of a QTL is indicated by a contiguous string of markers thatexhibit statistical correlation to the phenotypic trait. Thus it ispossible to indicate the location of the QTL and the presence or absenceof the QTL (and with that the phenotype) by other markers located withinthe QTL region.

The number of potentially useful markers is limited but may be verylarge, and a skilled person may easily identify additional markers tothose disclosed in the application. Any marker that is linked to theresistance as disclosed in the application can be used in markerassisted selection.

Thus, alternative markers can therefore be developed by methods known tothe skilled person and used to identify and select plants with an alleleor a set of alleles of a quantitative trait locus or loci according tothe present invention and as disclosed herein before.

For example, the nucleotide sequence of the amplification productobtained in PCR amplification using the primer pairs as indicated inTable 10 and Table 11, respectively, exhibiting a nucleotide sequence asgiven in SEQ ID NO: 1-12 and SEQ ID NOs: 13-26, can be obtained by thoseskilled in the art and new primers or primer pairs designed based on thenewly determined nucleotide sequence of the PCR amplification product.Furthermore, the markers according to the invention and disclosed hereinbefore could be positioned on a genetic map of pepper or other species,in particular Solanaceae species and known markers mapping in the sameor homolog or ortholog region(s) could be used as starting point fordeveloping new markers.

The nucleotide sequences of the amplification products obtained in PCRamplification using the primer pairs as indicated in Table 10 and Table11, respectively, exhibiting a nucleotide sequence as given in SEQ IDNO: 1-12 and SEQ ID NOs: 13-26, or part thereof can also be used ashybridization probes, for example to screen a BAC library, to identifyadditional linked nucleotide sequences.

Accordingly, the markers specifically disclosed in the present inventionmay also be used in the identification and/or development of new oradditional markers associated with the QTL of interest, which in turncan then be used in marker assisted breeding and/or the search ofrecombinants flanking the QTL, and/or fine-mapping, and/or cloning ofthe QTL.

There are several methods or approaches available, known to thoseskilled in the art, which can be used to identify and/or develop markersin linkage disequilibrium and/or linked to and/or located in the QTLregion, as well as markers that represent the actual causal mutationsunderlying the QTL. Without being fully exhaustive some approaches,known by those skilled in the art, include:

-   -   use of disclosed sequences/markers in hybridization approaches        to identify other sequence in the region of interest: primer        sequences as disclosed herein and/or marker/gene sequences (or        part thereof) that can be determined using the primer sequences        as disclosed herein may be used as (hybridization) probes in        isolating nucleic acid sequences/genes flanking the markers        and/or linked and/or associated and/or specific for the QTL        region from a genomic nucleic acid sample and/or RNA or cDNA        sample or pool of samples (for example screening of genomic        resources like BAC libraries or gDNA or cDNA library screening).    -   use of disclosed sequences/markers in PCR approaches to identify        other sequence in the region of interest: primer sequences as        disclosed herein and/or marker/-(candidate) gene sequences (or        part thereof) that can be determined using the primer sequences        as disclosed may be used as (PCR) amplification primers to        amplify a nucleic acid sequence/gene flanking and/or linked to        and/or associated with and/or specific for the QTL region from a        genomic nucleic acid sample and/or RNA or cDNA sample or pool of        samples either or not isolated from a specific plant tissue        and/or after specific treatment of the plant and from capsicum        or in principal any other organism with sufficient homology.    -   use of disclosed sequences/markers in PCR approaches to identify        other sequence in the region of interest: the nucleotide        sequences/genes of one or more markers can be determined after        internal primers for said marker sequences may be designed and        used to further determine additional flanking sequence/genes        within the QTL region and/or genetically linked and/or        associated with the trait.    -   use of disclosed sequences/markers in mapping and/or comparative        mapping approaches to identify markers in the same region(s)        (positioning of QTL on other maps): based on positional        information and/or marker information as disclosed herein,        markers, of any type, may be identified by genetic mapping        approaches, eventually (if already needed) by positioning of the        disclosed markers (by genetic mapping or extrapolation based on        common markers across maps) on a (high density) genetic map(s),        and/or integrated genetic or consensus map(s). Markers already        known and/or new markers genetically linked and/or positioned in        the vicinity of the disclosed markers and/or QTL region may be        identified and/or obtained and eventually used in OIL        (fine-)mapping and/or QTL cloning and/or MAS breeding        applications.    -   use of disclosed sequences/markers in ‘in-siloco’ approaches to        identify additional sequences/markers/(candidate)genes in QTL        region(s): primer sequences as disclosed herein and/or        marker/(candidate)gene sequences (or part thereof) that can be        determined using the primer sequences as disclosed herein or        based on linked markers may be used in ‘in-silico’ methods to        search sequence or protein databases (e.g. BLAST) for        (additional) flanking and/or homolog sequences/genes and/or        allelic diversity (both genomic and/or cDNA sequences or even        proteins and both originating from capsicum and/or any other        organism) genetically linked and/or associated with the traits        as described herein and/or located in the QTL region.    -   use of disclosed sequences/markers in physical mapping        approaches (positioning of QTL on physical map or genome        sequence): primer sequences as disclosed herein and/or        marker/gene sequences (or part thereof) that can be determined        using the primer sequences as disclosed herein or using other        markers genetically linked to the markers disclosed herein        and/or located in the QTL region may be positioned on a physical        map and/or (whole) genome sequence in principal of any organism        with sufficient homology to identify (candidate)        sequences/markers/genes applicable in QTL (fine-mapping) and/or        QTL cloning and/or MAS breeding applications.    -   use of disclosed sequences/markers to position QTL on other        (physical) maps or genomes (across species . . . for pepper        other Solanaceae as tomato and potato are of first interest of        course but model species like Arabidopsis may be used): primer        sequences as disclosed herein and/or marker/gene sequences (or        part thereof) that can be determined using the primer sequences        as disclosed herein may be used in comparative genome or        syntheny mapping approaches to identify homolog region and        homolog and/or ortholog sequences/(candidate)genes genetically        linked and/or positioned in the QTL region and applicable in QTL        (fine-mapping) and/or QTL cloning and/or MAS breeding        applications.    -   use of disclosed sequences/markers to select the appropriate        individuals allowing the identification of markers in region of        interest by genetic approaches: primer sequences and/or markers        as disclosed herein may be used to select individuals with        different/contrasting QTL alleles which in for example in        genetic association approaches and/or bulk segregant analysis        (BSA, Michelmore et al., 1991) can be used to identify        markers/genes in the specific region (QTL region) of interest        and/or associated or genetically linked to the described traits.    -   use of disclosed information to search for (positional)        candidate genes: the disclosed information may be used to        identify positional and/or functional candidate genes which may        be associated with the described traits and/or genetically        linked.

In one embodiment, the invention therefore relates to a cultivatedCapsicum annuum plant comprising a genome comprising at least one QTLwhich contributes to Bemisia resistance, which QTL is located onchromosome 3, wherein said at least one QTL can be identified by amolecular marker that exhibit statistical correlation to the phenotypictrait, which marker can be developed from a DNA segment containing saidQTL by methods known in the art, which segment is obtainable from aplant which has the genetic background of line 061M4387, particularlyfrom a plant which has the genetic background or architecture at the QTLof line 061M4387, but especially from a plant of line 061M4387,representative seed of which is deposited at NCIMB under Accession No.NCIMB 41428, or from a progeny or an ancestor thereof comprising saidQTL, and defined by at least one marker locus, particularly to at leasttwo marker loci, more particularly to at least three marker loci andeven more particularly to at least four marker loci, but especially toat least five and up to six marker loci, which marker loci are onchromosome 3 and co-segregate with the Bemisia resistance trait and canbe identified by a PCR oligonucleotide primer or a pair of PCRoligonucleotide primers selected from the group of primer pair 1represented by a forward primer of SEQ ID NO: 1 and a reverse primer ofSEQ ID NO: 2, identifying marker locus 1; primer pair 2 represented by aforward primer of SEQ ID NO: 3 and a reverse primer of SEQ ID NO: 4,identifying marker locus 2; primer pair 3 represented by a forwardprimer of SEQ ID NO: 5 and a reverse primer of SEQ ID NO: 6, identifyingmarker locus 3; primer pair 4 represented by a forward primer of SEQ IDNO: 7 and a reverse primer of SEQ ID NO: 8, identifying marker locus 4;primer pair 5 represented by a forward primer of SEQ ID NO: 9 and areverse primer of SEQ ID NO: 10, identifying marker locus 5; and primerpair 6 represented by a forward primer of SEQ ID NO: 11 and a reverseprimer of SEQ ID NO: 12, identifying marker locus 6.

In one embodiment, the invention therefore relates to a cultivatedCapsicum annuum plant comprising a genome comprising at least two QTLwhich contribute to Bemisia resistance, which QTL are located onchromosome 3 and 5, wherein said at least two QTL can be identified by amolecular marker that exhibit statistical correlation to the phenotypictrait, which marker can be developed from a DNA segment containing saidQTL by methods known in the art, which segment is obtainable from aplant which has the genetic background of line 061M4387, particularlyfrom a plant which has the genetic background or architecture at the QTLof line 061M4387, but especially from a plant of line 061M4387,representative seed of which is deposited at NCIMB under Accession No.NCIMB 41428, or from a progeny or an ancestor thereof comprising saidQTL, wherein a first QTL is located on chromosome 3 in the donor plantand genetically linked to at least one marker locus, particularly to atleast two marker loci, particularly to at least three marker loci andparticularly to at least four marker loci, particularly to at least fivemarker loci, particularly to at least six marker loci, which marker lociare on chromosome 3 and co-segregate with the Bemisia resistance traitand can be identified by a pair of PCR oligonucleotide primers 1 to 6 asgiven in SEQ ID NOs: 1 to 12 and wherein a second QTL is located onchromosome 5 in the donor plant and genetically linked to at least onemarker locus, particularly to at least two marker loci, particularly toat least three marker loci and particularly to at least four markerloci, particularly to at least five marker loci, particularly to atleast six marker loci, and up to seven marker loci, which marker lociare on chromosome 5 and co-segregate with the Bemisia resistance traitand can be identified by a pair of PCR oligonucleotide primers selectedfrom the group of primer pairs 7 to 13 as given in SEQ ID NOs: 13 to 26.

In one embodiment, the invention relates to a cultivated Capsicum annuumplant comprising a genome comprising at least one QTL which contributesto thrips resistance, which QTL is located on chromosome 5, wherein saidat least one QTL can be identified by a molecular marker that exhibitstatistical correlation to the phenotypic trait, which marker can bedeveloped from a DNA segment containing said QTL by methods known in theart, which segment is obtainable from a plant which has the geneticbackground of line 061M4387, particularly from a plant which has thegenetic background or architecture at the QTL of line 061M4387, butespecially from a plant of line 061M4387, representative seed of whichis deposited at NCIMB under Accession No. NCIMB 41428, or from a progenyor an ancestor thereof comprising said QTL, and defined by at least onemarker locus, particularly to at least two marker loci, moreparticularly to at least three marker loci and even more particularly toat least four marker loci, but especially to at least five and up to sixmarker loci, which marker loci are on chromosome 3 and co-segregate withthe Bemisia resistance trait and can be identified by a PCRoligonucleotide primer or a pair of PCR oligonucleotide primers selectedfrom the group of primer pair 7 represented by a forward primer of SEQID NO: 13 and a reverse primer of SEQ ID NO: 14, identifying markerlocus 7; primer pair 8 represented by a forward primer of SEQ ID NO: 15and a reverse primer of SEQ ID NO: 16, identifying marker locus 8;primer pair 9 represented by a forward primer of SEQ ID NO: 17 and areverse primer of SEQ ID NO: 18, identifying marker locus 9; primer pair10 represented by a forward primer of SEQ ID NO: 19 and a reverse primerof SEQ ID NO: 20, identifying marker locus 10; primer pair 11represented by a forward primer of SEQ ID NO: 21 and a reverse primer ofSEQ ID NO: 22, identifying marker locus 11; primer pair 12 representedby a forward primer of SEQ ID NO: 23 and a reverse primer of SEQ ID NO:24, identifying marker locus 12, and primer pair 13 represented by aforward primer of SEQ ID NO: 25 and a reverse primer of SEQ ID NO: 26,identifying marker locus 13.

In one embodiment, the invention therefore relates to a cultivatedCapsicum annuum plant comprising a genome comprising at least two QTLwhich contribute to Bemisia and thrips resistance, which QTL are locatedon chromosomes 3 and 5 and wherein said at least two QTL can beidentified by a molecular marker that exhibit statistical correlation tothe phenotypic trait, which marker can be developed from a DNA segmentcontaining said QTL by methods known in the art, which segment isobtainable from a plant which has the genetic background of line061M4387, particularly from a plant which has the genetic background orarchitecture at the QTL of line 061M4387, but especially from a plant ofline 061M4387, representative seed of which is deposited at NCIMB underAccession No. NCIMB 41428, or from a progeny or an ancestor thereofcomprising said QTL, wherein a first QTL is located on chromosome 3 inthe donor plant and genetically linked to at least one marker locus,particularly to at least two marker loci, particularly to at least threemarker loci and particularly to at least four marker loci, particularlyto at least five marker loci, particularly to at least six marker loci,which marker loci are on chromosome 3 and co-segregate with the Bemisiaresistance trait and can be identified by a pair of PCR oligonucleotideprimers 1 to 6 as given in SEQ ID NOs: 1 to 12 and a second QTL islocated on chromosome 5 in the donor plant and genetically linked to atleast one marker locus, particularly to at least two marker loci,particularly to at least three marker loci and particularly to at leastfour marker loci, particularly to at least five marker loci,particularly to at least six marker loci, and up to seven marker loci,which marker loci are on chromosome 5 and co-segregate with the Bemisiaand/or thrips resistance trait and can be identified by a pair of PCRoligonucleotide primers selected from the group of primer pairs 7 to 13as given in SEQ ID NOs: 13 to 26.

The markers according to the present invention may be used inmarker-assisted-selection and/or any other methods wherein plants havingor have not the QTL are traced. The markers may be either trans, or cismarkers. A trans marker indicates a polymorphism resulting fromintrogression of exogenous (donor) DNA into a recipient plant's genome,which polymorphism is linked in cis with the recipient genome, i.e.linked with the opposite allele. Thus, cis markers are linked with theallele of interest (favorable QTL allele from the donor), while transmarkers are linked with the opposite allele (from the recipient).

To determine the utility of the inbred line and its potential togenetically contribute to the hybrid progeny a test-cross is made withanother inbred line, and the resulting progeny phenotypically evaluated.Traits that may be recorded commonly involve traits that are related tofruit shape and fruit characteristics such as pointed or non pointedfruit, pungent or non pungent, red, yellow or orange. Plantcharacteristics as length of internodes, growing power and ramificationsare also considered together with specific virus resistances such as TMV(Tobacco Mosaic virus) and TSWV (Tomato Spotted wilt virus).

For genotyping, QTL mapping or association mapping DNA is extracted fromsuitable plant material such as, for example, leaf tissue. Inparticular, bulks of leaves of a plurality of plants are collected. DNAsamples are genotyped using a plurality of polymorphic SSR's, SNPs orany other suitable marker-type covering the entire pepper genome.

Joint-analysis of genotypic and phenotypic data can be performed usingstandard software such as, for example, the software QTLCartographer andPlabQTL. Plant introductions and germplasm can be screened for thealleles at the corresponding QTLs disclosed in Table 10 and Table 11,respectively, based on the nucleotide sequence(s) of the marker(s) atthe marker locus/loci linked to said QTL or any other marker known to belocated on chromosome 3 and chromosome 5, respectively, and themolecular weight of the allele(s) using one or more of the techniquesdisclosed herein or known to those skilled in the art.

The nucleic acid sequence of markers disclosed, linked markers or theQTL of the present invention may be determined by methods known to theskilled person. For example, a nucleic acid sequence comprising said QTLor a resistance-conferring part thereof may be isolated from aBemisia/Thrips resistant donor plant by fragmenting the genome of saidplant and selecting those fragments harbouring one or more markersindicative of said QTL. Subsequently, or alternatively, the markersequences (or parts thereof) indicative of said QTL may be used as (PCR)amplification primers, in order to amplify (a) nucleic acid sequence(s)comprising said QTL form a genomic nucleic acid sample or a genomefragment obtained from said plant. The nucleotide sequence of the QTL,and/or of any additional marker comprised therein, may be obtained bystandard sequencing methods.

The present invention therefore also relates to an isolated nucleic acid(preferably DNA but not limited to DNA) sequence that comprises a QTL ofthe present invention, or a Bemisia/Thrips resistance-conferring partthereof. Thus the markers discloses may be used for the identificationand isolation of one or more markers or genes from pepper or othervegetable crops, particularly Solanaceous crops that are linked orencode Bemisia/Thrips resistance.

The nucleotide sequence of additional linked markers or the QTL of thepresent invention may for instance also be resolved by determining thenucleotide sequence of one or more markers associated with the QTL anddesigning primers for said marker sequences that may then be used tofurther determine the sequence outside of said marker sequence. Forexample the nucleotide sequence of the SSR markers disclosed herein orany other markers predicted in the QTL region and/or linked to the QTLmay be obtained by sequencing the PCR amplification product of saidmarkers, well known in the art. Or alternatively using the markersequences in a PCR or as hybridization probes to identify linkednucleotide sequences by for example but not limited BAC screening.

EXAMPLES

The following Examples provide illustrative embodiments. In light of thepresent disclosure and the general level of skill in the art, those ofskill will appreciate that the following Examples are intended to beexemplary only and that numerous changes, modifications, and alterationscan be employed without departing from the scope of the presentlyclaimed subject matter.

Example 1 Breeding History Pepper Breeding Line 061M4387

Using a quantitative bioassay as described below in Example 2 incombination with the appropriate growing conditions a wild Capsicumannuum accession was identified as a source of resistance to Bemisiatabaci and to thrips infestations. Seeds of this wild accession withaccession no: CGN16975 and accession name: AC 1979 was obtained from theinstituut voor de Veredeling van Tuinbouwgewassen (now Centre forGenetic Resources), Wageningen, Netherlands. A population segregatingfor the Bemisia and thrips resistance was created by crossing this donorpepper plant with a susceptible recipient pepper plant. A segregatingpopulation consisting of a total of 333 DH lines was created for theidentification of one or more QTLs contributing to the Bemisia andthrips resistance.

The initial source of resistance wild accession CGN16975 was crossedwith a Dutch inbred of Syngenta selected in Westland (The Netherlands).The F3 progeny of this cross was identified as resistant, particularlyintermediately resistant, to white fly (visual observation) in Almeria(Spain).

A (BC1) cross was then generated between the F3 mentioned in theprevious paragraph and a Syngenta line developed in Almeria. The progenyof this cross was identified as resistant, particularly intermediatelyresistant, to white fly and thrips (in Agadir-Morocco; thrips—visualobservations) and used for the next cross (BC2).

The BC2 was generated with a Syngenta dihaploid line developed inAlmeria (Spain). From plant families in the F2 of this cross several(=333) dihaploids were developed to characterize the heredity of thetrait of resistance to white fly and thrips. Among them, line 061M4387,deposited with NCIMB under accession no. NCIMB 41428 was shown to beintermediately resistant to Bemisia tabaci and thrips infestations.

Example 2 Resistance Assay

2A Bemisia Resistance—Testing Protocol

2A.1 Plant Raise

Plants were Sown and Raised According to Standard Procedures

In particular, plants were sown in a e.g. 77 multitray (multipots of4.2×4.2×5.9 cm) filled with well drained, friable soil with a pH between6.5 to 7.5 and grown for approx. one month in a tunnel. The plants werefertilized with N 4.0-6.0 P 0.35-1.0, K 4.0-6.0 of EC2.

The plants were transplanted according to special design (see below) andcultivated according to standard procedures. During plant culture,plants often need to be protected against parasites other than Bemisia.Chemical treatment was carried out with pesticides that only have aminor impact on the population development of Bemisia.

2A.2 Insect Culture and Inoculation

In order to ensure a stable and uniform development of the Bemisiapopulation and to obtain more stringent testing conditions, an earlyinoculation of the plants with Bemisia was carried out. For this aseparate small plastic greenhouse was used in which a pepper crop wasraised under standard conditions. The naturally present Bemisiapopulation was used to inoculate the test material.

Two Methods can be Used for Inoculation:

-   1) Use of squash as a trap plant 2-3 week old seedlings of squash    were placed in the small plastic greenhouse with the Bemisia culture    for 4-6 hours. Because Bemisia adults have a strong preference for    squash, they will rapidly fly to the squash seedlings. The squash    plants with the Bemisia adults were then carefully enveloped with a    plastic bag and transferred to the experimental tunnel and    homogeneously released over the plants. Inoculation starts ca.10d    after transplantation and can be continued for 1-2 wk as necessary.-   2) The young pepper seedlings raised in trays were placed 4-5 days    before transplantation in the small plastic greenhouse with the    Bemisia culture allowing the adult Bemisia to lay eggs on the young    plants. Hereafter the plants can be transplanted to the larger    bitunnel.    2A3 Experimental Design

A large bi-tunnel in Agadir was used with 8 rows of ca. 2×150 plantseach. In each row one side was used as spreader row and planted with asusceptible entry (existing F1 e.g. Bikingo F1 or Vergasa F1, or,susceptible parental line). The other part of the row was planted withthe test entries. The test entries were fully randomized in each of atleast 7 blocks with 1 or more plants/entry per block.

2A.4 Data Collection

Bemisia development was monitored in the spreader row weekly or biweeklyby assessing 5 spreader row plants on equidistant positions (plant 1,38, 75, 112, 150) in each row. The final assessments were made when theaverage infestation of the monitored spreader plants was approx. 4(Table 1) usually at the time when the first fruits are ripening (3-4months after transplantation).

For assessing the severity, a scale from 1-9 was used (Table 1). Theabaxial side of the leaves of the plant was inspected and the average ofthe ca. 5 worst affected leaves was assessed according the 1-9 scale.All test plants were scored in this way.

TABLE 1 Assessments scale for WF-resistance. scale description WF ##pupae¹ 9 no pupae 0 8 very few pupae 1-5 7 some pupae irregularscattered over leaf  5-20 6 20-50 5 moderate number of pupae moreregular distributed  50-100 over leaves 4 100-200 3 many pupae, denselycrowded on leaf (black mould 200-400 present) 2 400-700 1 plantcompletely covered with pupae and heavily  700-1000 moulded oftenstunted in growth ¹estimation of ## pupae per leaf: empty pupal casesand mature pupae2A.5 Data Analysis

Data were analyzed by calculating the means per test entry andcomparison with a susceptible entry (e.g., susceptible spreader row orelse). A multiple comparison of the means (e.g. LSD) indicates if testentries differ mutually and from a susceptible control.

2A.6 Results

2A.6.1 Resistance Testing

Table 2 shows some of the results of the Bemisia screening in Agadir(Morocco) of the deposited line 061M438 7 (containing QTL1 (referred toherein as the QTL on chromosome 3) and QTL2 (referred to herein as theQTL on chromosome 5), see example 3), two of its ancestors and theresistance donor. The breeding history is explained in example 1. Thedeposited line 061M4387 proved to be significantly better under varyinginsect pressures, seasons and conditions compared to standardsusceptible varieties or lines.

TABLE 2 Results of tests with deposited line 061M4387 and some of itsancestors. Entry Resistant line Susc. control¹ test remark CGN16975(donor) very resistant* *scored visually in BC1F3 very resistant* verysusc.* spring 2002 breeding trial Mean n Mean LSD² CGN16975 (donor) 8.820 6.6 0.4 December 2002 insect pressure BC1F4 8.5 20 relatively lowdeposited line Mean n Mean LSD 061M4387 8.0 7 4.2 1.2 August 2005moderate-high insect pressure 061M4387 6.4 7 3.0 1.5 November 2005 veryhigh insect pressure 061M4387 6.7 20 3.8 0.6 June 2006 high insectpressure 061M4387 8.0 10 5.2 1.4 February 2007 moderate insect pressure¹In the above trials Vergasa F1, Bikingo F1 or parental lines (allsusceptible) are used as both susceptible control and as spreader row.²LSD intervals(P < 0.05) are based on comparison of means of manyentries included in trial (not shown).2A.6.2 QTL Identification Associated with Resistance

Table 3 shows the results of a screening of a set of 333 DH linesdeveloped out of a BC2F2 for the purpose of identification of QTLsassociated with resistance (see examples 1 and 3). Two tests wereperformed on the DH-lines: test 1 was planted in spring 2005 and scoredin August, the second test was planted in September 2005 and scored inNovember. The average infestation in August was lower compared toNovember (Table 3). In the November trial some spots in the greenhousehad a very high insect pressure with as consequence that no reliablephenotypes could be obtained. Those spots were therefore excluded fromthe analysis.

Two QTLs were identified (see example 3). The QTL (QTL1) on chromosome 3had the largest LOD-value (up to 50), the QTL (QTL2) on chromosome 5 hada smaller LOD-value (up to 5).

Based on flanking markers 69 DH-lines were identified having no QTLs, 57DH-lines had QTL1 only, 38 DH-lines had QTL2 only, and, 63 DH-linespossessed QTL1 and QTL2 together.

The effect of QTL1 was in both trials significant and estimated in theAugust trial 1.3 scale units and in November trial 1.9 scale units(Table 3). Due to the lower infestation in August many lines scored a7-8 reaching a plafond possibly minimizing differences between lines. Inthe November trial the differences between lines were larger due to thehigher infestation level.

The effect of QTL2 was more pronounced in the August trial (0.6 scaleunit) compared to the November trial.

TABLE 3 Results of a screening of a set of 333 DH lines developed out ofa BC2F2 for the purpose of identification of QTLs associated withresistance no QTL QTL1 QTL2 QTL1&2 August 5.9a* 7.2c 6.5b 7.6d November4.1a 6.0b 4.3a 6.1b Avg 5.0 6.6 5.4 6.8 *similar letters show nostatistical difference (LSD, P < 0.05) within observation period246.3 Test of Effect of QTL1 in 5 BC3 Families

To estimate the effect of QTL1 in different background families, fiveBC3's were made with 5 different BC-parents and a derivative resistantline containing QTL1. 556 DH-lines derived from these BC3's weregenotyped for QTL1 and tested in Agadir (February 2007, Table 4) underthe same conditions described previously. The effect of QTL1 wasestimated between 1.8-2.3 scale units for the different families (Table4) confirming the significant effect of QTL1 (Two-way ANOVA, Mean SquareQTL1=521.3, F=698.7, P<0.001, no interactions between family and QTL1presence).

TABLE 4 Effect of QTL1 on Bemisia infestation in 5 BC3 families testedin Agadir February 2007. family QTL1 presence mean n diff. 1 no QTL 5.251 1.8 QTL1 7.0 46 2 no QTL 4.7 47 2.0 QTL1 6.8 38 3 no QTL 5.2 60 2.1QTL1 7.3 145 4 no QTL 4.9 37 2.2 QTL1 7.2 57 5 no QTL 4.9 42 2.3 QTL17.3 33 total 55624.6.4 Test of the Effect of QTLs 1 and 2 on Bemisia Damage in a Set of60 DH Lines Derived from a Cross with Donor CGN16975 and Two ElitePepper Lines.

The initial source of resistance wild accession CGN16975 was crossedwith two elite lines (A and B). From these two F1's in total 60dihaploid lines (34 from line A and 26 from line B) were derived. 58 ofthese lines were tested mostly twice (between December 2003 and December2004) for Bemisia resistance.

TABLE 5 Estimated effect of QTL 1 and 2 on Bemisia resistance in a smallDH population derived out of F1 between wild accession CGN16975 and 2elite pepper lines. no QTL QTL1 QTL2 QTL1&2 n = 11 n = 17 n = 17 n = 13average 4.0a* 5.9.0b 5.1ab 5.9b Given is the average of 2 tests.*Similar letters indicate no significant differences (ANOVA, followed bycomparison of means with Fisher's Least Significance Difference methodLSD, P < 0.05). **significantly different from no QTL at P = 0.06.

The effect of QTL1 was estimated on ca. 1.9 scale units and the effectof QTL 2 was estimated on 1.1 scale units in these populations.

2B Thrips Resistance—Testing Protocol

2B.1 Plant Raise

Plants were sown in standard peat soil and transplanted after 14d into7×7×8× cm pots. The plants were grown in a greenhouse at 20° C./18° C.and 16 hr/8 hr day/night. Approximately 1 month after sowing, the plantswere transferred to a 1×1×1 m cage covered with a nylon mesh (0.07×0.27mm) preventing thrips from leaving the cage. In each cage 400-500 thripswere released. This was repeated 1 week later to ensure a high insectpressure. Three-four weeks after the first inoculation, the observationwas done.

2B.2 Insect Culture and Inoculation

A viable culture of thrips was maintained and used for resistanceexperiments.

From the culture 400-500 thrips (including both adults and juveniles)were collected in a vial with a standard insect sucking device. The vialwith thrips was subsequently released in the test cage. Afterinoculation the temperature was raised to 24° C. continuous (day/night).

28.3 Experimental Design

In each cage one or more resistant controls (preferably CGN16975) andone or more susceptible controls (e.g. Roxy F1 and/or Snooker F1) wereplaced. In total 57 plants can be placed in a single cage. The plants(including the control plants) were randomized for each cage. The DHlines tested for Bemisia (see Example 1 above) were tested for thripsresistance in this way. Seven consecutive experiments were performed inorder to test all 333 DH lines with (max.) 12 plants per line,

28.4 Data Collection

The final assessments were made when the average infestation of thesusceptible control plants was 3-4 (Table 6). Usually this is reached3-4 weeks after inoculation.

For assessing silvering damage, a scale from 1-9 was used (Table 6). Theabaxial side of the leaves of the plant was inspected and the average ofthe ca. 2-3 worst affected leaves was assessed according the 1-9 scale.All test plants were scored in this way.

TABLE 6 Assessment scale for silvering damage caused by Frankliniellaoccidentalis. scale description Thrips damage % silvering¹ 9 nosilvering damage 0 8 tiny spots <0.1 7 some small spots especially nearthe mid vein or 0.1-1   edge of the leaf 6 1-2 5 moderate number ofspots more regular distributed 3-5 over leaves 4  6-10 3 many largesilvering spots present distributed 11-20 over the entire leaf 2 21-40 1very heavy silvering, large part of the leaf damaged >40 ¹estimation of% silvering of 2-3 most affected leaves2B.5 Data Analysis

Data were analyzed by calculating the means per test entry andcomparison with a susceptible entry (e.g. Roxy F1 and/or Snooker F1). Amultiple comparison of the means (e.g. LSD) indicated if test entriesdiffer mutually and from a susceptible control.

2B.6 Results

2B6.1 Resistance Testing

Table 7 shows as an example the results of CGN16975, two susceptiblecontrols (Roxy F1 and Snooker F1) and the deposited line 061M4387possessing QTL1 and QTL2. Averages of 3 independent tests each with ca.12 plants/entry are given. The deposited line showed significantly lesssilvering compared to the two susceptible control lines Snooker F1 andRoxy F1 but more silvering compared to the donor CGN16975. Thisindicates that the deposited line has an elevated level of resistancecompared to standard varieties.

TABLE 7 Phenotyping results of deposited line 061M4387, CGN16975 and twosusceptible varieties (Roxy F1 and Snooker F1). entry n silvering*Snooker F1 36 3.5a Roxy F1 36 3.8a 061M4387 31 5.8b CGN16975 36 7.3c*Simlar letters: indicate no significant differences (ANOVA, followed bycomparison of means with Fisher's Least Significance Difference methodLSD, P < 0.05).28.6.2 QTL Identification Associated with Resistance to Thrips

The QTL-analysis (see example 3) on the 333 DH lines (see example 1)revealed a QTL on chromosome 5, with a LOD value up to 12. This QTL waslocated in the same region as QTL2 identified in the Bemisia QTL mapping(see example 2A.6)

The QTL for Bemisia on chromosome 5 and the QTL for thrips are locatedon the same chromosomal region. It could be a single QTL with an effectboth against Bemisia and thrips or two linked QTLs

28.6.3 Test of Effect of QTL2 on Thrips Damage in a Set of 333 DH LinesDeveloped Out of a BC2F2

The effect of the thrips QTL was estimated similarly as was done forBemisia (see 2A.6) based on the large 333 DH set. The QTL showed asignificant effect of ca. 0.8 scale unit (Table 8).

TABLE 8 Estimated effect of QTL 2 on thrips damage (silvering) in largeBC2F2 population of 333 DH lines (possible recombinants were excluded).no QTL QTL2 n = 122 n = 91 Silvering 4.5 5.3* *Significant difference(t-test, t = −8.29, P < 0.001).28.6.4 Test of the Effect of QTL2 on Thrips Damage in a Set of 60 DHLines Derived from a Cross with Donor CGN16975 and Two Elite PepperLines.

53 DH lines of the same set of 60 DH lines derived from accessionCGN16975 crossed with two elite lines (A and B, see section 2A.6.4) weresubjected twice to a thrips test and checked for the presence of QTL2.

QTI2 showed a significant effect of 1.0 scale units (Table 9) which ison a comparable level as in the large 333 DH set described in 2B.6.3(Table 8).

TABLE 9 Estimated effect of QTL 2 on thrips damage (silvering) in asmall DH population derived out of F1 between wild accession CGN16975and 2 elite pepper lines. no QTL QTL2 n = 26 n = 27 Silvering 4.3 5.3**Significant difference (t-test, t = −3.86, P < 0.001).

Example 3 QTL Mapping

3.1 QTL Mapping for Bemisia Resistance

Using the quantitative bioassay described above in Example 2A incombination with appropriate growing conditions a source of resistanceto Bemisia was identified. A population segregating for the Bemisiaresistance was created by crossing this donor pepper plant with asusceptible recipient pepper plant. A segregating population consistingof a total of 333 DH lines was created for the identification of QTLcontributing to the Bemisia resistance. DNA was extracted from a pool ofleaves of 8 individual plants of each DH line and the parent plants ofthe population following standard protocols. The parents of thepopulation were screened using several hundred SSR's in order toidentify SSR's which are polymorphic between the parents. Subsequentlythe DH population was genotyped using the identified polymorphic SSRmarkers. Based on the so obtained segregation data a molecular markermap was prepared using the commonly used software Mapmaker and Joinmap.The markers represent genome regions polymorphic between the parents ofthe population.

QTL mapping, i.e. joint analysis of genotypic and phenotypic data wasperformed using the QTLCartographer software. QTLs were identified whichare located on different chromosomes including a QTL on chromosome 3,which was demonstrated to be associated to Bemisia resistance. The QTLis characterized by means of markers positioned on the genetic map andmarker alleles of markers known to be located in the QTL region. Therebythe location of a/multiple resistance conferring DNA sequences is/areestablished. Details of the QTL associated with resistance to Bemisia,i.e. flanking markers and markers located in the QTL region arerepresented in Table 10.

3.2 QTL Mapping for Thrips Resistance

The identical approach as described in Example 3.1 above was taken formapping the QTL associated with resistance to thrips.

A QTL, which was demonstrated to be associated to thrips resistance, wasidentified on chromosome 5. The QTL is characterized by means of markerspositioned on the genetic map and marker alleles of markers known to belocated in the OIL region. Thereby the location of a/multiple resistanceconferring DNA sequences is/are established. Details of the QTLassociated with resistance to thrips, i.e. flanking markers and markerslocated in the QTL region are represented in Table 11.

TABLE 10 Details of the QTL associated with resistance to Bemisia, i.e.flanking markers and markers located in the QTL region Bemisia F PrimerR Primer Resistance Chromosome Marker Linked Sequence ID Sequence ID QTL# # Locus Marker Forward Primer Number Reverse Primer Number 1 3 1 LM1001 TGCTGGGAAAGATCT SEQ.ID.NO: 1 ATCAAGGAAGCAAACCA SEQ.ID.NO: 2 CAAAAGGATGC 1 3 2 LM 1002 GCAGCGTTACCAAAT SEQ.ID.NO: 3 TGTTTGCTATTCAATATASEQ.ID.NO: 4 AACCG TGCTTTGA 1 3 3 LM 1003 GGAAGCTTAGCCACA SEQ.ID.NO: 5ACCATATTTCCGACTTTG SEQ.ID.NO: 6 CATC AAC 1 3 4 LM 1004 TCCATCATCGACTGGSEQ.ID.NO: 7 TGTTCAATTGGCTTCTGT SEQ.ID.NO: 8 AGAC G 1 3 5 LM 1005GCAAGTAGAACAAAG SEQ.ID.NO: 9 TATTTGAAGGTTGTGCG SEQ.ID.NO: 10 GGTAGG AC 13 6 LM 1006 TCATCACATTCACTT SEQ.ID.NO: 11 TTGATTCATTTCAGATAG SEQ.ID.NO:12 CATTTTC TTCAAG

TABLE 11 Details of the QTL associated with resistance to thrips, i.e.flanking markers and markers located in the QTL region Bemisia/ Thrips FPrimer R Primer Resistance Chromosome Marker Linked Sequence ID SequenceID QTL # # Locus Marker Forward Primer Number Reverse Primer Number 2 5 7 LM 2001 CTTTGGAGGTAGCG SEQ.ID.NO: 13 CAACAAACGAACCACAA SEQ.ID.NO: 14GTATG TG 2 5  8 LM 2002 CCCGTTTACAAGCA SEQ.ID.NO: 15 GACCCCTGAAGAACCTCSEQ.ID.NO: 16 AAGAG TC 2 5  9 LM 2003 TCTCTTGTCAGACA SEQ.ID.NO: 17CTTCTTGGAGGCATTTTT SEQ.ID.NO: 18 CGTCG G 2 5 10 LM 2004 TGTAGGATTACAAGSEQ.ID NO: 19 GCGAGCTATTACACCGA SEQ.ID.NO: 20 AACATTATCG AG 2 5 11 LM2005 TAGGTGGGAATACA SEQ.ID.NO: 21 CCCAGATCTACCAAGGA SEQ.ID.NO: 22 CTGGGGTC 2 5 12 LM 2006 TCGGCCTGACTAGT SEQ.ID.NO: 23 CGGGTACCAGATGTAGGSEQ.ID.NO: 24 ATTGAC G 2  5? 13 LM 2007 ATCGTGAGGTGAGT SEQ.ID.NO: 25TACCTACATACCCCCACC SEQ.ID.NO: 26 ACGAG C

DEPOSIT

Applicants have made a deposit with an effective date of 10 Aug. 2006 ofat least 2500 seeds of Capsicum annuum line 061M4387 with the NCIMB Ltd,Ferguson Building, Craibstone Estate, Bucksburn, Aberdeen, AB21 9YA,Scotland, under accession no: NCIMB 41428.

The foregoing invention has been described in detail by way ofillustration and example for purposes of clarity and understanding.However, it will be obvious that certain changes and modifications suchas single gene modifications and mutations, somaclonal variants, variantindividuals selected from large populations of the plants of the instantinbred and the like may be practiced within the scope of the invention,as limited only by the scope of the appended claims. Thus, although theforegoing invention has been described in some detail in this document,it will be obvious that changes and modification may be practiced withinthe scope of the invention, as limited only by the scope of the appendedclaims.

All references cited herein are incorporated by reference in theapplication in their entireties.

The invention claimed is:
 1. A method for introducing at least oneallele associated with resistance to Bemisia at a quantitative traitlocus (“QTL”) contributing to resistance to Bemisia into a Capsicumannuum plant lacking said allele comprising: a) obtaining a firstCapsicum annuum plant wherein said plant contains a genome comprising atleast one QTL located on chromosome 3 or chromosome 5 which contributesto Bemisia resistance; b) crossing said first Capsicum annuum plant witha second Capsicum annuum plant, wherein said second Capsicum annuumplant lacks said QTL; and c) identifying a C. annuum plant resultingfrom the cross exhibiting increased resistance to Bemisia comprisingdetecting, by using at least one oligonucleotide primer or primer pair,at least one marker allele located on chromosome 3 or chromosome 5 andco-segregating with said Bemisia resistance, wherein said at least onemarker allele co-segregating with said Bemisia resistance ischaracterized by the PCR amplification product of i. a PCR primer orprimer pair selected from the group consisting of primer pairs 1-6represented by forward and reverse primers of SEQ ID NOs: 1-12,including primer pairs resulting from a combination of the forward andreverse primers of SEQ ID NOs: 1-12; and/or ii. a PCR primer pairselected from the group consisting of primer pairs 7-13 represented byforward and reverse primers of SEQ ID NOs: 13-26, including primer pairsresulting from a combination of the forward and reverse primers of SEQID NOs: 13-26.
 2. A method for introducing at least a first allele at aquantitative trait locus (“QTL”) contributing to resistance to Bemisiaand at least a second allele at a QTL contributing to resistance tothrips, into a Capsicum annuum plant lacking said alleles comprising: a)obtaining a first plant of the genus Capsicum, particularly a Capsicumannuum plant wherein said plant contains a genome comprising a firstallele at a QTL located on chromosome 3 or chromosome 5 contributing toresistance to Bemisia and at least a second allele at a QTL located onchromosome 5 contributing to resistance to thrips; b) crossing saidfirst plant of the genus Capsicum, particularly a Capsicum annuum plant,with a second Capsicum annuum plant, wherein said second Capsicum annuumplant lacks said QTL; and c) identifying a plant resulting from thecross exhibiting increased resistance to Bemisia and thrips, comprisingdetecting, by using at least one oligonucleotide primer or primer pair,at least one marker allele co-segregating with the Bemisia resistanceand at least one marker allele co-segregating with the thripsresistance, wherein said at least one marker allele co-segregating withsaid Bemisia resistance is located on chromosome 3 or chromosome 5 andis characterized by the PCR amplification product of i. a PCR primer orprimer pair selected from the group consisting of primer pairs 1-6represented by forward and reverse primers of SEQ ID NOs: 1-12,including primer pairs resulting from a combination of the forward andreverse primers of SEQ ID NOs: 1-12; and/or ii. a PCR primer or primerpair selected from the group consisting of primer pairs 7-13 representedby forward and reverse primers of SEQ ID NOs: 13-26, including primerpairs resulting from a combination of the forward and reverse primers ofSEQ ID NOs: 13-26; and wherein said at least one marker alleleco-segregating with said thrips resistance is located on chromosome 5and is characterized by the PCR amplification product of a PCR primer orprimer pair selected from the group consisting of primer pairs 7-13represented by forward and reverse primers of SEQ ID NOs: 13-26,including primer pairs resulting from a combination of the forward andreverse primers of SEQ ID NOs: 13-26.
 3. A method for introducing atleast one allele associated with resistance to Bemisia at a quantitativetrait locus (“QTL”) contributing to resistance to Bemisia into aCapsicum annuum plant lacking said allele comprising: a) obtaining afirst Capsicum annuum plant wherein said plant contains a genomecomprising at least one QTL located on chromosome 3 which contributes toBemisia resistance; b) crossing said first Capsicum annuum plant with asecond Capsicum annuum plant, wherein said second Capsicum annuum plantlacks said QTL; and c) identifying a C. annuum plant resulting from thecross exhibiting increased resistance to Bemisia comprising detecting,by using an oligonucleotide primer or primer pair, at least one markerallele located on chromosome 3 and co-segregating with said Bemisiaresistance, wherein said at least one QTL is obtained from a plant ofline 061 M4387, representative seed of which is deposited underAccession No. NCIMB 41428, or from a progeny or ancestor thereofcomprising said QTL.
 4. A method for introducing at least one alleleassociated with resistance to Bemisia at a quantitative trait locus(“QTL”) contributing to resistance to Bemisia into a Capsicum annuumplant lacking said allele comprising: a) obtaining a first Capsicumannuum plant wherein said plant contains a genome comprising at leastone QTL located on chromosome 3 which contributes to Bemisia resistance;b) crossing said first Capsicum annuum plant with a second Capsicumannuum plant, wherein said second Capsicum annuum plant lacks said QTL;and c) identifying a C. annuum plant resulting from the cross exhibitingincreased resistance to Bemisia comprising detecting, by using at leastone oligonucleotide primer or primer pair, at least one marker alleleco-segregating with said Bemisia resistance, wherein said first Capsicumannuum plant is a plant of line 061 M4387, representative seed of whichis deposited under Accession No. NCIMB 41428, or a progeny thereof. 5.The method of claim 1, wherein the oligonucleotide primer or primer pairis selected from the group consisting of primer pairs 1-6 represented byforward and reverse primers of SEQ ID NOs: 1-12, including primer pairsresulting from a combination of the forward and reverse primers of SEQID NOs: 1-12.
 6. The method of claim 1, wherein the oligonucleotideprimer or primer pair is selected from the group consisting of primerpairs 7-13 represented by forward and reverse primers of SEQ ID NOs:13-26, including primer pairs resulting from a combination of theforward and reverse primers of SEQ ID NOs: 13-26.